Article

Expression of heat-shock protein Hsp60 correlated with the apoptotic index and patient prognosis in human oesophageal squamous cell carcinoma

January 2005
European Journal of Cancer 40(18):2804-11

January 2005
40(18):2804-11

DOI:10.1016/j.ejca.2004.08.013

Source
PubMed

Authors:

Ahmad Faried

Universitas Padjadjaran

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Cellular stress response and apoptosis are two highly conserved mechanisms for maintaining homeostasis. Hsp60 and Hsp90 have been shown to play pro- and anti-apoptotic roles, respectively. Our present study examined whether there is a correlation between the expression of Hsp60 and Hsp90, clinical parameters, the apoptotic index (AI), and the prognosis of patients with oesophageal squamous cell carcinoma (ESCC). We immunohistochemically stained cells for Hsp60, Hsp90, and single-stranded DNA (ssDNA), which acts as an apoptotic marker. In normal oesophageal epithelium tissue, Hsp60 and Hsp90 were expressed in the cytoplasm and membrane from the basal cell layer to the supra-basal cell layers. Hsp60 and Hsp90 positive stainings (+) were found in 63 of 123 cases (51%) and 62 of 123 cases (50%), respectively. There was no correlation between Hsp60 and Hsp90 expression levels and any of the clinical parameters examined. The five-year survival rate for ESCC patients with Hsp60 (+) expression was significantly higher than for those patients with Hsp60 (-) expression (P=0.0371). Five-year survival rates of patients with Hsp60 (+) and (-) were 49% and 33%, respectively. By contrast, Hsp90 expression failed to predict patient prognosis (P=0.7965). The high-AI group did not have a significantly better prognosis than the low-AI group (P=0.2218). Statistical analysis showed a significant correlation between the expression of Hsp60 and AI in ESCC patients (P=0.008). Thus, the five-year survival rate for the high-AI/Hsp60 (+) group was statistically significantly better than for the other groups (P=0.0281). The results obtained in this study indicate that positive Hsp60 expression is a good prognostic indicator. This may be due to its role as a chaperone in contributing to the induction of apoptosis. These data suggest that Hsp60 expression correlates with the AI and patient prognosis in human ESCC.

The Role of the Heat-Shock Proteins in Esophagogastric Cancer

Article

Full-text available

Aug 2022

Heat-shock proteins (HSPs) are a family of proteins that have received considerable attention over the last several years. They have been classified into six prominent families: high-molecular-mass HSP, 90, 70, 60, 40, and small heat shock proteins. HSPs participate in protein folding, stability, and maturation of several proteins during stress, such as in heat, oxidative stress, fever, and inflammation. Due to the immunogenic host’s role in the combat against cancer cells and the role of the inflammation in the cancer control or progression, abnormal expression of these proteins has been associated with many types of cancer, including esophagogastric cancer. This study aims to review all the evidence concerning the role of HSPs in the pathogenesis and prognosis of esophagogastric cancer and their potential role in future treatment options. This narrative review gathers scientific evidence concerning HSPs in relation to esophagus and gastric cancer. All esophagogastric cancer subtypes are included. The role of HSPs in carcinogenesis, prognostication, and therapy for esophagogastric cancer are discussed. The main topics covered are premalignant conditions for gastric cancer atrophic gastritis, Barrett esophagus, and some viral infections such as human papillomavirus (HPV) and Epstein–Barr virus (EBV). HSPs represent new perspectives on the development, prognostication, and treatment of esophagogastric cancer.

HSP60 in cancer: a promising biomarker for diagnosis and a potentially useful target for treatment

Article

Full-text available

May 2021

Heat shock proteins (HSPs), most of which are molecular chaperones, are highly conserved proteins produced by cells under physiological stress or pathological conditions. HSP60 (57–69 kDa) can promote or inhibit cell apoptosis through different mechanisms, and its abnormal expression is also related to tumour cell metastasis and drug resistance. In recent years, HSP60 has received increasing attention in the field of cancer research due to its potential as a diagnostic and prognostic biomarker or therapeutic target. However, in different types of cancer, the specific mechanisms of abnormally expressed HSP60 in tumour carcinogenesis and drug resistance are complicated and still require further study. In this article, we comprehensively review the regulative mechanisms of HSP60 on apoptosis, its applications as a cancer diagnostic biomarker and a therapeutic target, evidence of involvement in tumour resistance and the applications of exosomal HSP60 in liquid biopsy. By evaluating the current findings of HSP60 in cancer research, we highlight some core issues that need to be addressed for the use of HSP60 as a diagnostic or prognostic biomarker and therapeutic target in certain types of cancer.

Prognosis value of heat-shock proteins in esophageal and esophagogastric cancer: A systematic review and meta-analysis

Article

Full-text available

Apr 2024

BACKGROUND Heat shock proteins (HSPs) are molecular chaperones that play an important role in cellular protection against stress events and have been reported to be overexpressed in many cancers. The prognostic significance of HSPs and their regulatory factors, such as heat shock factor 1 (HSF1) and CHIP, are poorly understood. AIM To investigate the relationship between HSP expression and prognosis in esophageal and esophagogastric cancer. METHODS A systematic review was conducted in accordance with PRISMA recommendations (PROSPERO: CRD42022370653), on Embase, PubMed, Cochrane, and LILACS. Cohort, case-control, and cross-sectional studies of patients with esophagus or esophagogastric cancer were included. HSP-positive patients were compared with HSP-negative, and the endpoints analyzed were lymph node metastasis, tumor depth, distant metastasis, and overall survival (OS). HSPs were stratified according to the HSP family, and the summary risk difference (RD) was calculated using a random-effect model. RESULTS The final selection comprised 27 studies, including esophageal squamous cell carcinoma (21), esophagogastric adenocarcinoma (5), and mixed neoplasms (1). The pooled sample size was 3465 patients. HSP40 and 60 were associated with a higher 3-year OS [HSP40: RD = 0.22; 95% confidence interval (CI): 0.09-0.35; HSP60: RD = 0.33; 95%CI: 0.17-0.50], while HSF1 was associated with a poor 3-year OS (RD = -0.22; 95%CI: -0.32 to -0.12). The other HSP families were not associated with long-term survival. HSF1 was associated with a higher probability of lymph node metastasis (RD = -0.16; 95%CI: -0.29 to -0.04). HSP40 was associated with a lower probability of lymph node dissemination (RD = 0.18; 95%CI: 0.03-0.33). The expression of other HSP families was not significantly related to tumor depth and lymph node or distant metastasis. CONCLUSION The expression levels of certain families of HSP, such as HSP40 and 60 and HSF1, are associated with long-term survival and lymph node dissemination in patients with esophageal and esophagogastric cancer.

Function and Fiber-Type Specific Distribution of Hsp60 and αB-Crystallin in Skeletal Muscles: Role of Physical Exercise

Article

Full-text available

Jan 2021

Skeletal muscle is a plastic and complex tissue, rich in proteins that are subject to continuous rearrangements. Skeletal muscle homeostasis can be affected by different types of stresses, including physical activity, a physiological stressor able to stimulate a robust increase in different heat shock proteins (HSPs). The modulation of these proteins appears to be fundamental in facilitating the cellular remodeling processes related to the phenomenon of training adaptations such as hypertrophy, increased oxidative capacity, and mitochondrial activity. Among the HSPs, a special attention needs to be devoted to Hsp60 and αB-crystallin (CRYAB), proteins constitutively expressed in the skeletal muscle, where their specific features could be highly relevant in understanding the impact of different volumes of training regimes on myofiber types and in explaining the complex picture of exercise-induced mechanical strain and damaging conditions on fiber population. This knowledge could lead to a better personalization of training protocols with an optimal non-harmful workload in populations of individuals with different needs and healthy status. Here, we introduce for the first time to the reader these peculiar HSPs from the perspective of exercise response, highlighting the control of their expression, biological function, and specific distribution within skeletal muscle fiber-types.

Heat Shock Protein 60 Overexpression Is Associated with the Progression and Prognosis in Gastric Cancer

Article

Full-text available

Sep 2014
PLOS ONE

Heat shock protein 60 (HSP60) is a chaperonin with essential functions for cell physiology and survival, and its expression correlates with prognosis in a number of malignancies. The aim of this study is to determine the relationship of HSP60 status with clinicopathological parameters and prognosis in gastric cancer.The levels of HSP60 and matrix metallopeptidase 9 (MMP-9) antigen was evaluated by immunohistochemistry in 223 gastric carcinoma samples. The association between HSP60 and MMP-9, clinicopathological parameters, and prognosis of gastric cancer was examined.The level of HSP60 protein was significantly associated with depth invasion, lymph node metastasis and stage of disease (all P

Heat Shock Proteins as Prognostic Markers of Cancer

Article

Sep 2014

This review systematically examines the literature and data on the prognostic significance of heat shock proteins (heat shock protein - Hsp) Hsp27, Hsp60, Hsp70, Hsp90 in various cancers. Our analysis of the literature showed that the existing data are contradictory with regard to the prognostic significance of Hsp. This result may be due to biological differences of the carcinomas studied and methodological differences in the assessment of heat shock proteins. Heat shock protein is removed a significant role in the development of cancer. In cancer heat shock proteins, are expressed in large amounts, allow tumor cells to escape apoptotic death. Therefore, the potential scheme of inhibiting Hsp is currently therapeutically attractive direction against cancer.

Heat Shock Protein 60 Overexpression Is Associated with the Progression and Prognosis in Gastric Cancer

Article

Full-text available

May 2014
J CANCER RES CLIN

The heat shock protein 22 (HSP22) is associated with tumor proliferation and protects tumor cell from apoptosis in many malignancies. However, the role of HSP22 in gastric cancer has not been thoroughly elucidated. The aim was to determine the relationship of HSP22 expression with clinicopathological parameters and prognosis in gastric cancer and estimate the alteration of HSP22 expression after neoadjuvant chemotherapy. HSP22 and matrix metallopeptidase 9 (MMP-9) antigen expressions were evaluated by immunohistochemistry in 129 gastric carcinoma samples. Univariate and multivariate analyses were performed to determine the association between HSP22 expression and prognosis. The response of HSP22 was assessed in 47 patients who received neoadjuvant chemotherapy. HSP22 protein expression was significantly associated with tumor size, depth invasion, lymph node metastasis and stage of disease (all P < 0.05). In univariate and multivariate analyses, HSP22 was an independent prognostic factor for both overall survival (OS) and recurrence-free survival (RFS) (P = 0.003 and P = 0.004, respectively). Furthermore, HSP22 overexpression was associated with a poor prognosis in all patients and in patients subgroups stratified by tumor size, depth invasion and lymph node metastasis. In addition, HSP22 was significantly correlated with MMP-9 among 129 gastric cancer tissues (P < 0.001). Patients who had MMP-9 overexpression had poor OS and shorter RFS. Moreover, the alteration of HSP22 was not comparable in 47 patients who underwent neoadjuvant chemotherapy. HSP22 plays an important role on tumor aggressiveness and prognosis and may act as a promising target for prognostic prediction.

Polydatin and Nicotinamide Rescue the Cellular Phenotype of Mitochondrial Diseases by Mitochondrial Unfolded Protein Response (mtUPR) Activation

Article

Full-text available

May 2024

Primary mitochondrial diseases result from mutations in nuclear DNA (nDNA) or mito-chondrial DNA (mtDNA) genes, encoding proteins crucial for mitochondrial structure or function. Given that few disease-specific therapies are available for mitochondrial diseases, novel treatments to reverse mitochondrial dysfunction are necessary. In this work, we explored new therapeutic options in mitochondrial diseases using fibroblasts and induced neurons derived from patients with mutations in the GFM1 gene. This gene encodes the essential mitochondrial translation elongation factor G1 involved in mitochondrial protein synthesis. Due to the severe mitochondrial defect, mutant GFM1 fibroblasts cannot survive in galactose medium, making them an ideal screening model to test the effectiveness of pharmacological compounds. We found that the combination of polydatin and nicotinamide enabled the survival of mutant GFM1 fibroblasts in stress medium. We also demonstrated that polydatin and nicotinamide upregulated the mitochondrial Unfolded Protein Response (mtUPR), especially the SIRT3 pathway. Activation of mtUPR partially restored mitochondrial protein synthesis and expression, as well as improved cellular bioenergetics. Furthermore, we confirmed the positive effect of the treatment in GFM1 mutant induced neurons obtained by direct reprogramming from patient fibroblasts. Overall, we provide compelling evidence that mtUPR activation is a promising therapeutic strategy for GFM1 mutations.

Mechanistic study of heat shock protein 60-mediated apoptosis in DF-1 cells

Article

Full-text available

Mar 2024
POULTRY SCI

Heat shock proteins (HSP) are a group of highly conserved molecular chaperones found in various organisms and have been associated with tumorigenesis, tumor progression, and metastasis. However, the relationship between HSP60 and apoptosis remains elusive. The aim of this study was to explore the role and regulatory mechanisms of apoptosis in response to altered HSP60 expression. We generated DF-1 cell lines of both HSP60 overexpression and knockdown and assessed their impact on apoptosis levels using ELISA and flow cytometry analyses. Additionally, we examined the transcription and protein expression levels of apoptosis-related signaling factors using fluorescence quantitative PCR (qPCR) and Western blotting analyses. Heat shock proteins 60 overexpression led to a significant decrease in apoptosis levels in DF-1 cells, which could be attributed to the downregulation of BAX and BAK expression, the upregulation of Bcl-2, and the decreased expression of Caspase 3. Conversely, HSP60 knockdown led to a substantial increase in apoptosis levels in DF-1 cells, facilitated by the downregulation of BAX and Bcl-2 expression, and the upregulation of BAK expression, which increased Caspase 3 levels, thereby promoting apoptosis. The findings of our study provide the first evidence of the inhibitory effect of HSP60 on apoptosis in DF-1 cells. These observations have significant implications for disease progression and cancer research, with potential medical applications.

High expression of HSP60 and survivin predicts poor prognosis for oral squamous cell carcinoma patients

Article

Full-text available

Sep 2023
BMC Oral Health

Background HSP60 is a heat shock proteins (HSPs) family member and help mitochondrial protein to fold correctly. Survivin is one of the inhibitors of apoptosis protein family member, which plays a significant part in cancer progression. They were capable of forming HSP60-survivin complexes and involved in the development of various tumors. Methods The Cancer Genome Atlas (TCGA) database demonstrated that HSP60 and survivin and their correlation on mRNA expression level with OSCC patients. Besides, expression of HSP60 and survivin proteins was studied utilizing immunohistochemistry in tissue microarrays (TMA) in OSCC and in adjacent non-cancerous squamous epithelium (Non-CCSE) tissues. Results Significantly increased levels of HSP60 and survivin in most cancers compared to normal tissue by pan-cancer analysis. HSP60 and survivin proved a significantly increased expression in OSCC samples compared to Non-CCSE both on mRNA and protein (both P < 0.05). Additionally, elevated HSP60 displayed a positive correlation with survivin in terms of mRNA and protein expression levels (all P < 0.001). Patients with OSCC who had advanced clinical stage or lymph node metastasis (LNM) showed higher HSP60 expression (P = 0.004, P = 0.006, respectively). Higher levels of the proteins HSP60 and survivin were significantly inversely correlated relationship with OSCC patients’ overall survival rates in multivariate survival analysis (P = 0.018, P = 0.040). From the above results, overexpression of HSP60 and survivin protein may serve as independent biomarkers predicting poor prognosis in OSCC. Conclusions Elevated HSP60 and survivin might be served as novel poor prognosis biomarkers for surgically resected OSCC patients.

Clinicopathologic Significance of Heat Shock Protein 60 as a Survival Predictor in Colorectal Cancer

Article

Full-text available

Aug 2023

Simple Summary HSP60, a mitochondrial chaperone, can promote or inhibit cancer progression. Patients with colorectal cancer (CRC) were examined for HSP60 expression using the TNM classification. Patients with differentiated and p53-mutated CRC expressed high levels of HSP60. Compared with patients with high HSP60 expression, those with low expression had event-free and disease-specific survival hazard ratios of 1.42 and 1.69, respectively. TNM class and HSP60 expression affected survival, especially in late/advanced stages. The expression of the HSPD1 gene, which encodes HSP60, exhibited the same pattern as the protein. The hazard ratios for overall and relapse-free survival were 1.80 and 1.87, respectively, for patients with reduced HSPD1 expression. Low HSPD1 expression and advanced malignancy worsen CRC prognosis. This study suggests that HSPD1/HSP60 may be a useful biomarker for refined survival prediction in late-stage and advanced-stage CRC, allowing for individualized therapy. Abstract The role of heat shock protein 60 (HSP60), a mitochondrial chaperone, in tumor progression or its anti-tumor effects remains controversial. This study aimed to confirm the possibility of using HSP60 as a prognostic marker in patients with colorectal cancer (CRC), considering TNM classification for precise prediction. HSP60 expression increased with differentiation and p53 mutations in patients. However, compared to patients with high HSP60 expression, patients with low HSP60 expression had event-free survival and disease-specific survival hazard ratios (HRs) of 1.42 and 1.69, respectively. Moreover, when the survival rate was analyzed by combining TNM classification and HSP60 expression, the prognosis was poor, particularly when HSP60 expression was low in the late/advanced stage. This pattern was also observed with HSP family D member 1, HSPD1, the gene that encodes HSP60. Low HSPD1 expression was linked to lower overall survival and relapse-free survival rates, with HRs of 1.80 and 1.87, respectively. When TNM classification and HSPD1 expression were considered, CRC patients with low HSPD1 expression and advanced malignancy had a poorer prognosis than those with high HSPD1 expression. Thus, HSPD1/HSP60 can be a useful biomarker for a sophisticated survival prediction in late- and advanced-stage CRC, allowing the design of individualized treatment strategies.

Knockdown of heat shock protein family D member 1 (HSPD1) promotes proliferation and migration of ovarian cancer cells via disrupting the stability of mitochondrial 3-oxoacyl-ACP synthase (OXSM)

Article

Full-text available

Apr 2023

Background Heat shock protein 60 (HSP60) is essential for the folding and assembly of newly imported proteins to the mitochondria. HSP60 is overexpressed in most types of cancer, but its association with ovarian cancer is still in dispute. SKOV3 and OVCAR3 were used as experimental models after comparing the expression level of mitochondrial HSP60 in a normal human ovarian epithelial cell line and four ovarian cancer cell lines. Results Low HSPD1 (Heat Shock Protein Family D (HSP60) Member 1) expression was associated with unfavorable prognosis in ovarian cancer patients. Knockdown of HSPD1 significantly promoted the proliferation and migration of ovarian cancer cells. The differentially expressed proteins after HSPD1 knockdown were enriched in the lipoic acid (LA) biosynthesis and metabolism pathway, in which mitochondrial 3-oxoacyl-ACP synthase (OXSM) was the most downregulated protein and responsible for lipoic acid synthesis. HSP60 interacted with OXSM and overexpression of OXSM or LA treatment could reverse proliferation promotion mediated by HSPD1 knockdown. Conclusions HSP60 interacted with OXSM and maintained its stability. Knockdown of HSPD1 could promote the proliferation and migration of SKOV3 and OVCAR3 via lowering the protein level of OXSM and LA synthesis.

High Expression of Heat Shock Protein Family D Member 1 Predicts Poor Prognosis of Esophageal Cancer

Article

Full-text available

Jul 2022

Background: Heat shock protein family D (Hsp60) member 1 (HSPD1) has been reported as a potential survival-related biomarker in some cancers. However, the correlation between HSPD1 expression with prognosis and clinical features of esophageal cancer (EC) is poorly understood. Our research aimed to explore the clinical and prognostic significance of HSPD1 expression in EC patients. Methods: In our study, HSPD1 expression was detected by immunochemistry in 87 EC tissue specimens and 20 normal cancerous peripheral tissue specimens. Meanwhile, we also analyzed the expression of HSPD1 in EC by The Cancer Genome Atlas (TCGA) database. Then Chi-squared and Fisher's exact tests and Wilcoxon signed-rank test and logistic regression models were separately used to test the correlation between clinical characteristics and HSPD1 expression in our and TCGA cohort. Moreover, we evaluated the value of HSPD1 in prognosis by Kaplan-Meier curves and Cox analysis. Finally, gene set enrichment analysis (GSEA) was performed using the data accessed from TCGA. Results: The results showed that HSPD1 was overexpressed in EC, and the expression was related to histological type, histological grade, N classification, and clinical stage. Moreover, Kaplan-Meier curves and Cox analysis indicated that high expression of HSPD1 correlated with poor prognosis, and HSPD1 was an independent risk factor for EC. GSEA identified pathways involved in cysteine and methionine metabolism, spliceosome, selenoamino acid metabolism, mismatch repair, RNA degration, DNA replication, and cell cycle as differentially enriched in ECs with high HSPD1 expression. Conclusions: Our results suggest that HSPD1 is expressed at high levels in EC, and has potential to be used as a novel biomarker for the prognosis of patients with EC.

The mitochondrial unfolded protein response (UPR): shielding against toxicity to mitochondria in cancer

Article

Full-text available

Jul 2022
J HEMATOL ONCOL

Mitochondria are essential for tumor growth and progression. However, the heavy demand for mitochondrial activity in cancer leads to increased production of mitochondrial reactive oxygen species (mtROS), accumulation of mutations in mitochondrial DNA, and development of mitochondrial dysfunction. If left unchecked, excessive mtROS can damage and unfold proteins in the mitochondria to an extent that becomes lethal to the tumor. Cellular systems have evolved to combat mtROS and alleviate mitochondrial stress through a quality control mechanism called the mitochondrial unfolded protein response (UPR mt ). The UPR mt system is composed of chaperones and proteases, which promote protein folding or eliminate mitochondrial proteins damaged by mtROS, respectively. UPR mt is conserved and activated in cancer in response to mitochondrial stress to maintain mitochondrial integrity and support tumor growth. In this review, we discuss how mitochondria become dysfunctional in cancer and highlight the tumor-promoting functions of key components of the UPR mt .

Heat shock proteins with an emphasis on HSP 60

Article

Full-text available

Oct 2021
MOL BIOL REP

Heat shock phenomenon is a process by which cells express a set of proteins called heat shock proteins (HSPs) against heat stress. HSPs include several families depending upon the molecular weight of the respective protein. Among the different HSPs, The HSP60 is one of the main components representing the framework of chaperone system. HSP60 plays a myriad number of roles like chaperoning, thermotolerance, apoptosis, cancer, immunology and embryonic development. In this review we discussed briefly the general knowledge and focussed on HSP60 in terms of structure, regulation and function in various physiological and pathological conditions.

Heat shock protein 60 and cardiovascular diseases: An intricate love‐hate story

Article

Full-text available

Aug 2020

Cardiovascular diseases (CVDs) are the result of complex pathophysiological processes in the tissues comprising the heart and blood vessels. Inflammation is the main culprit for the development of cardiovascular dysfunction, and it may be traced to cellular stress events including apoptosis, oxidative and shear stress, and cellular and humoral immune responses, all of which impair the system's structure and function. An intracellular chaperone, heat shock protein 60 (HSP60) is an intriguing example of a protein that may both be an ally and a foe for cardiovascular homeostasis; on one hand providing protection against cellular injury, and on the other triggering damaging responses through innate and adaptive immunity. In this review we will discuss the functions of HSP60 and its effects on cells and the immune system regulation, only to later address its implications in the development and progression of CVD. Lastly, we summarize the outcome of various studies targeting HSP60 as a potential therapeutic strategy for cardiovascular and other diseases.

Heat Shock Protein 60 in Hepatocellular Carcinoma: Insights and Perspectives

Article

Full-text available

Apr 2020

Heat shock protein 60 (HSP60) is a mitochondrial chaperone that is implicated in physiological and pathological processes. For instance, it contributes to protein folding and stability, translocation of mitochondrial proteins, and apoptosis. Variations in the expression levels of HSP60 have been correlated to various diseases and cancers, including hepatocellular carcinoma (HCC). Unlike other HSPs which clearly increase in some cancers, data about HSP60 levels in HCC are controversial and difficult to interpret. In the current review, we summarize and simplify the current knowledge about the role of HSP60 in HCC. In addition, we highlight the possibility of its targeting, using chemical compounds and/or genetic tools for treatment of HCC.

Functional Compartmentalization of HSP60-Survivin Interaction between Mitochondria and Cytosol in Cancer Cells

Article

Full-text available

Dec 2019

Heat shock protein 60 (HSP60) and survivin reside in both the cytosolic and mitochondrial compartments under physiological conditions. They can form HSP60-survivin complexes through protein-protein interactions. Their expression levels in cancer tissues are positively correlated and higher expression of either protein is associated with poor clinical prognosis. The subcellular location of HSP60-survivin complex in either the cytosol or mitochondria is cell type-dependent, while the biological significance of HSP60-survivin interaction remains elusive. Current knowledge indicates that the function of HSP60 partly rests on where HSP60-survivin interaction takes place. HSP60 has a pro-survival function when binding to survivin in the mitochondria through interacting with other factors such as CCAR2 and p53. In response to cell death signals, mitochondrial survivin functions through preventing procaspase activation. Degradation of cytosolic survivin leads to the loss of mitochondrial membrane potential and aberrant mitosis processes. On the other hand, HSP60 release from mitochondria to cytosol upon death stimuli might exert a pro-death function, either through stabilizing Bax, enhancing procaspase-3 activation, or increasing protein ubiquitination. Combining the knowledge of mitochondrial HSP60-survivin complex function, cytosolic survivin degradation effect, and pro-death function upon mitochondria release of HSP60, a hypothetical scenario for HSP60-survivin shuttling upon death stimuli is proposed.

Heat Shock Proteins 27 and 60 Serum Levels in Patients with Gastrointestinal Cancer and Acute Myocardial Infarction in Birjand, Iran

Article

Full-text available

Jun 2018
IMMINV

Introduction: Cancer and myocardial infarction are lethal diseases. Their prevalence is increasing worldwide. In both diseases, the level of oxidative stress rises because of tissue damage. The aim of this study was to evaluate the serum levels of heat shock protein 27 and heat shock protein 60 in patients with cancer and myocardial infarction, and then compare them with healthy individuals. Materials and Methods: After blood samples were collected from the participants, plasma and serum were separated from these samples for further examination. The serum levels of heat shock protein 27 and heat shock protein 60 were measured with related kits in 30 patients with cancer and 30 patients with acute myocardial infarction, followed by 30 healthy individuals. The collected data were then analyzed in the Statistical Package for the Social Sciences software (version 22). Results: The mean serum levels of heat shock protein 27 in cancer patients (25.21 ± 5.57 ng/mL) and in patients with myocardial infarction (45.23 ± 7.43) were significantly higher than those in healthy individuals (10.61 ± 3.11; P

S1 Table

Data

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Apr 2018

Primers used for qRT-PCR. (PDF)

Targeting HSP60 by subcutaneous injections of jetPEI/HSP60‐shRNA destabilizes cytoplasmic survivin and inhibits hepatocellular carcinoma growth

Article

Sep 2017

Heat shock protein 60 (HSP60) overexpresses in various types of cancer, but its expression levels and functions in hepatocellular carcinoma (HCC) are still in dispute. We aim to clarify this issue and examine whether HSP60 could be a therapeutic target for HCC. We found drastically enhanced cell apoptosis and suppressed cell proliferation in two HCC cell lines with HSP60‐silencing, and also indicated survivin is involved in this regulatory process in vitro and in vivo. However, HSP60‐silencing in normal human hepatocytes only resulted in a minimal reduction of cell proliferation but without effects on cell apoptosis. We also showed HSP60 interacted with cytosolic but not mitochondrial survivin by immunoprecipitation assay. A rigorous method was used to standardized quantitative from immunoblot assay to obtain more precise expression level of HSP60 and survivin. The expression of HSP60 and survivin positively correlated in both cancerous and non‐cancerous liver tissues (P < 0.001) after analyzing 145 surgically removed HCC tissues. 56.6% of HCC patients were overexpressed HSP60 in cancerous tissues, and 40.0% were under‐expressed HSP60. Higher expression of HSP60 and survivin in non‐cancerous tissues both correlated with shorter overall survival (P = 0.029 and P < 0.001, respectively). Finally, we evaluated the therapeutic potential of HSP60 using extraneous delivery of jetPEI/shHSP60 complexes. The treatment results showed the significant reduction of tumor weight by 44.3% (P < 0.05), accompanied by under‐expression of survivin. These studies suggested that HSP60 not only as a prognostic marker but also could serve as a novel therapeutic target for HCC. This article is protected by copyright. All rights reserved

Investigation of the Cell Stabilization and the Epithelial to Mesenchymal Transition Effect of Flavopiridol in Mouse Lung Squamous Cell Carcinoma

Article

Jan 2016

Abstract Background: Lung malignancy is a leading cause of cancer related morbidity and mortality worldwide. The three main subtypes of Non-small cell lung cancer (NSCLC) are adenocarcinoma, squamous cell lung cancers (SqCLCs) and large-cell carcinoma. Flavopiridol is a flavone that inhibits several cyclindependent kinases and exhibits potent growthinhibitory activity, apoptosis and G1 phase arrest in a number of human tumor cell lines. The present study focused on the effect of flavopiridol in cell stabilization, cell adhesion, junctional complex and epithelial to mesenchymal transition (EMT) in mouse lung squamous cell carcinoma cell. Methods: Cell viability and proliferation of untreated controls and flavopiridol treated cells were determined by using the WST-1 assay. SqCLCs and M. dunni mouse skin fibroblast cells (MSF) cells immunofluorescence analyses were performed for the evaluation of Hsp90β, e-cadherin and occludin. The percentages of CD133+/CD44+ cells in the flavopiridol treated cells when compared with the control untreated cells by flow cytometric analysis. Results: Flow cytometric analysis showed that the percentages of CD133+ and CD44+ cells did not show significant changes when compared to the untreated cells. Hsp90b expression which is significantly available in SqCLCs was considered as important its significant reduction by following flavopiridol application and also flavopiridol caused the significant increase the E-cadherin expression but there was no significant effect on the occludin expression. SqCLCs cells are uniformly highly sensitive to flavopiridol induced cytotoxicity during prolonged 72 h exposure to clinically achievable concentrations of this drug. Conclusions: These observations may have translational implications for the use of flavopiridol in the treatment of SqCLCs.

Investigation of the Cell Stabilization and the Epithelial to Mesenchymal Transition Effect of Flavopiridol in Mouse Lung Squamous Cell Carcinoma

Article

Full-text available

Jan 2015

Background: Lung malignancy is a leading cause of cancer related morbidity and mortality worldwide. The three main subtypes of Non-small cell lung cancer (NSCLC) are adenocarcinoma, squamous cell lung cancers (SqCLCs) and large-cell carcinoma. Flavopiridol is a flavone that inhibits several cyclindependent kinases and exhibits potent growthinhibitory activity, apoptosis and G1 phase arrest in a number of human tumor cell lines. The present study focused on the effect of flavopiridol in cell stabilization, cell adhesion, junctional complex and epithelial to mesenchymal transition (EMT) in mouse lung squamous cell carcinoma cell. Methods: Cell viability and proliferation of untreated controls and flavopiridol treated cells were determined by using the WST-1 assay. SqCLCs and M. dunni mouse skin fibroblast cells (MSF) cells immunofluorescence analyses were performed for the evaluation of Hsp90β, e-cadherin and occludin. The percentages of CD133+/CD44+ cells in the flavopiridol treated cells when compared with the control untreated cells by flow cytometric analysis. Results: Flow cytometric analysis showed that the percentages of CD133+ and CD44+ cells did not show significant changes when compared to the untreated cells. Hsp90b expression which is significantly available in SqCLCs was considered as important its significant reduction by following flavopiridol application and also flavopiridol caused the significant increase the E-cadherin expression but there was no significant effect on the occludin expression. SqCLCs cells are uniformly highly sensitive to flavopiridol induced cytotoxicity during prolonged 72 h exposure to clinically achievable concentrations of this drug. Conclusions: These observations may have translational implications for the use of flavopiridol in the treatment of SqCLCs

Neonatal Death and Heart Failure in Mouse with Transgenic HSP60 Expression

Article

Full-text available

Oct 2015
BMRI

Mitochondrial heat shock proteins, such as HSP60, are chaperones responsible for the folding, transport, and quality control of mitochondrial matrix proteins and are essential for maintaining life. Both prosurvival and proapoptotic roles have been proposed for HSP60, and HSP60 is reportedly involved in the initiation of autoimmune, metabolic, and cardiovascular diseases. The role of HSP60 in pathogenesis of these diseases remains unclear, partly because of the lack of mouse models expressing HSP60. In this study we generated HSP60 conditional transgenic mice suitable for investigating in vivo outcomes by expressing HSP60 at the targeted organ in disease models. Ubiquitous HSP60 induction in the embryonic stage caused neonatal death in mice at postnatal day 1. A high incidence of atrial septal defects was observed in HSP60-expressing mice, with increased apoptosis and myocyte degeneration that possibly contributed to massive hemorrhage and sponge-like cardiac muscles. Our results showed that neonatal heart failure through HSP60 induction likely involves developmental defects and excessive apoptosis. The conditional HSP60 mouse model is useful for studying crucial biological questions concerning HSP60.

Role of heat shock protein HSP90 and HSP70 in macrophagic differentiation

Article

May 2010

David Lanneau

Heat shock proteins (HSPs) are molecular chaperones whose expression is increased after many different stresses. They have a protective function helping the cell to cope with lethal conditions. These proteins play an essential role as molecular chaperones by assisting the correct folding of nascent and stress-accumulated misfolded proteins and by preventing their aggregation. My team is interested in understanding the roles of HSPs in two physiological related processes: apoptosis and cell differentiation. The aim of my work is to study the functions of HSP90 and HSP70 in macrophagic differentiation. I first studied the role of HSP90 in macrophagic differentiation. We previously reported that cellular inhibitor of apoptosis protein-1 (c-IAP1), migrated from the nucleus to the surface of the Golgi apparatus in cells undergoing differentiation. c-IAP1 is a member of the inhibitor of apoptosis protein (IAP) family which has demonstrated functions in cell death, cell signaling and mitosis. Here, we show that c-IAP1 is a client protein of the stress protein HSP90β. In three distinct cellular models, the two proteins interact and migrate from the nucleus to the cytoplasm during the differentiation process through a leptomycin B-sensitive pathway. Inhibition of HSP90 proteins by small chemical molecules and specific depletion of HSP90isoform by siRNA both leads to c-IAP1 degradation by the proteasomal machinery. This chaperone function of HSP90 towards c-IAP1 is specific of its β isoform as specific depletion of HSP90α isoform does not affect c-IAP1 content. Chemical inhibition of HSP90 or siRNA-mediated depletion of HSP90both inhibit cell differentiation, which can be reproduced by siRNA-mediated depletion of c-IAP1. Altogether, these results suggest that HSP90prevents auto-ubiquitination and degradation of its client protein c-IAP1, whose depletion would be sufficient to inhibit cell differentiation. The second part of my work consisted in the study of the role of HSP70 in macrophagic differentiation. We used two models of differentiation: human peripheral monocytes exposed to M-CSF and THP1 cells induced to differentiate by TPA. We found that in both models, HSP70 expression was induced during differentiation. Interestingly, the expression of Spi-1/Pu.1, a transcription factor essential for monocytes to differentiate, was similarly induced. Upon differentiation, both proteins co-localized in the nucleus and associated. Inhibition or down regulation of HSP70 induced Spi-1/Pu.1 degradation and blocks the differentiation process, indicating that the necessity of Spi-1/Pu.1 to be chaperoned by HSP70 during differentiation. Since Spi-1/Pu.1 promoter has a HSE-like, we studied whether transcription factors responsible for HSPs induction, HSF, could be involved. We show that although HSF2 do not seem involved, HSF1 binds to Spi-1/Pu.1 promoter and its inhibition blocks Spi-1/Pu.1 expression and monocytes differentiation. So HSP90 and HSP70 are essentials for macrophagic differentiation. Understanding monocyte differentiation regulation is important since defects of the differentiation process can lead to the development of leukemias (acute myeloid leukemia, chronic myelomonocytic leukemia). A better understanding of the roles of HSPs may provide new therapeutic strategies for the treatment of these pathologies.

Cytosolic Hsp60 Can Modulate Proteasome Activity in Yeast

Article

Full-text available

Dec 2014

Hsp60, an essential oligomeric molecular mitochondrial chaperone, has been subject to rigorous basic and clinical research. With yeast as a model system, we provide evidence for the ability of cytosolic yHsp60 to inhibit the yeast proteasome: i) Following biological turnover of Murine-Bax (a proteasome substrate), we show that co-expression of cytosolic yHsp60 stabilizes Bax, enhances its association with mitochondria and enhances its killing capacity. ii) Expression of yHsp60 in the yeast cytosol (yHsp60c) inhibits degradation of a cytosolic protein ΔMTS-Aco1 tagged with the degron SL17 (a ubiquitin-proteasome substrate). iii) Conditions under which Hsp60 accumulates in the cytosol (elevated Hsp60c or growth at 37oC) correlate with reduced 20S peptidase activity in proteasomes purified from cell extracts. iv) Elevated yHsp60 in the cytosol correlate with accumulation of poly-ubiquitinated proteins. v) According to 20S proteasome pull down experiments, Hsp60 is physically associated with proteasomes in extracts of cells expressing Hsp60c or grown at 37oC. Even mutant Hsp60 proteins, lacking chaperone activity, were still capable of proteasome inhibition. The results support the hypothesis, that localization of Hsp60 to the cytosol may modulate proteasome activity according to cell need. Copyright © 2014, The American Society for Biochemistry and Molecular Biology.

Combination of SNX-2112 with 5-FU exhibits antagonistic effect in esophageal cancer cells

Article

Full-text available

Oct 2014

The low efficacy of single-drug chemotherapy forms the basis for combination therapy in esophageal squamous cell carcinoma. SNX-2112, a selective heat shock protein 90 (Hsp90) inhibitor, was recently reported as being effective in combination with cisplatin and paclitaxel. In this study, we investigated the effect of SNX-2112 in combination with 5-fluorouracil (5-FU), another first-line anticancer drug, in esophageal cancer. Unexpectedly, tetrazolium assay revealed that the combination of SNX-2112 with 5-FU exhibited antagonistic effect. Flow cytometry revealed that the SNX-2112 and 5-FU combination greatly decreased the number of G2/M cells compared to SNX-2112-only treatment in Eca‑109 cells. This effect might be related to the altered mRNA level of cyclin-related genes including cyclin D1, Chk2 and Cdk4. Further, 5-FU attenuated SNX-2112-induced apoptosis by decreasing poly(ADP-ribose) polymerase (PARP) cleavage and inactivating caspase-3, -8 and -9. Additionally, 5-FU suppressed the SNX-2112-induced decrease of mitochondrial membrane potential. Moreover, 5-FU partly recovered Hsp90 client proteins, including Akt, p-Akt, inhibitor of κB kinase (IKK)α, extracellular signal-regulated kinase (ERK)1/2, and glycogen synthase kinase (GSK)-3β, which SNX-2112 had downregulated. Taken together, this is the first report that the combination of SNX-2112 with 5-FU exhibited antagonistic effect in esophageal cancer cells by affecting growth inhibition, cell cycle, apoptosis, and Hsp90 client proteins, suggesting that care is required in the clinical application of combined SNX-2112 and 5-FU.

Mitochondrial Transcription Factor A Worsens the Clinical Course of Patients With Pancreatic Cancer Through Inhibition of Apoptosis of Cancer Cells

Article

Apr 2014
PANCREAS

Mitochondrial transcription factor A (mtTFA) is mandatory for both the transcription and maintenance of mitochondrial DNA. This study aimed to investigate the significance of mtTFA expression in pancreatic ductal adenocarcinoma (PDAC). Surgical specimens from 93 patients with PDAC who all underwent pancreatectomy were immunohistochemically stained using a polyclonal anti-mtTFA antibody. The relationship between the expression of mtTFA, clinicopathologic factors, and prognosis of these patients were evaluated. Positive mtTFA expression was significantly associated with lymphovascular invasion and metastatic recurrence in the liver and correlated with an advanced surgical stage. A univariate analysis showed that the patients with positive mtTFA expression had a significantly shorter survival time than those patients with negative mtTFA expression, and a multivariate analysis revealed that mtTFA expression was one of the independent prognostic factors in patients with PDAC. Positive mtTFA expression was significantly correlated with a low apoptotic index but not significantly correlated with the mind bomb homolog-1 (MIB-1) index. The expression mtTFA worsens the clinical course of patients with PDAC through the inhibition of apoptosis of PDAC cells and is an independent marker for the poor prognosis of the patients with PDAC after pancreatectomy. Mitochondrial transcription factor A may be a novel target for the treatment of PDAC.

Hsp60 chaperonopathies and chaperonotherapy: Targets and agents

Article

Full-text available

Nov 2013
EXPERT OPIN THER TAR

Introduction: Hsp60 (Cpn60) assembles into a tetradecamer that interacts with the co-chaperonin Hsp10 (Cpn10) to assist client polypeptides to fold, but it also has other roles, including participation in pathogenic mechanisms. Area covered: Hsp60 chaperonopathies are pathological conditions, inherited or acquired, in which the chaperone plays a determinant etiologic-pathogenic role. These diseases justify selection of Hsp60 as a target for developing agents that interfere with its pathogenic effects. We provide information on how to proceed. Expert opinion: The information available encourages the development of ways to improve Hsp60 activity (positive chaperonotherapy) when deficient or to block it (negative chaperonotherapy) when pathogenic. Many questions are still unanswered and obstacles are obvious. More information is needed to establish when and why autologous Hsp60 becomes a pathogenic autoantigen, or induces cytokine formation and inflammation, or favors carcinogenesis. Clarification of these points will take considerable time. However, analysis of the Hsp60 molecule and a search for active compounds aimed at structural sites that will affect its functioning should continue without interruption. No doubt that some of these compounds will offer therapeutic hopes and will also be instrumental for dissecting structure-function relationships at the biochemical and biological (using animal models and cultured cells) levels.

Promising Urinary Protein Biomarkers for the Early Detection of Hepatocellular Carcinoma among High-Risk Hepatitis C Virus Egyptian Patients

Article

Full-text available

Sep 2012

Hepatocellular Carcinoma is a major healthcare problem, representing the third most common cause of cancer-related mortality worldwide. There are 130 million Hepatitis C virus infected patients worldwide who are at a high-risk for developing Hepatocellular Carcinoma. Due to the fact that reliable parameters and/or tools for the early detection of Hepatocellular Carcinoma among high-risk individuals are severely lacking, Hepatocellular Carcinoma patients are always diagnosed at a late stage where surgical solutions or effective treatment are not possible. Urine was collected from 106 Hepatitis C infected patients patients, 32 of whom had already developed Hepatocellular Carcinoma and 74 patients who were diagnosed as Hepatocellular Carcinoma -free at the time of initial sample collection. In addition to these patients, urine samples were also collected from 12 healthy control individuals. Total urinary proteins were isolated from the urine samples and LC-MS/MS was used to identify potential protein HCC biomarker candidates. This was followed by validating relative expression levels of proteins present in urine among all the patients using quantitative real time-PCR. This approach revealed that significant over-expression of three proteins: DJ-1, Chromatin Assembly Factor-1 (CAF-1) and Heat Shock Protein 60 (HSP60), was a characteristic event among Hepatocellular Carcinoma - post Hepatitis C virus infected patients. As a single-based Hepatocellular Carcinoma biomarker, CAF-1 over-expression identified Hepatocellular Carcinoma among Hepatitis C virus infected patients with a specificity of 90%, sensitivity of 66% and with an overall diagnostic accuracy of 78%. Moreover, the CAF-1/HSP60 tandem identified Hepatocellular Carcinoma among Hepatitis C virus infected patients with a specificity of 92%, sensitivity of 61% and with an overall diagnostic accuracy of 77%.

Mitochondrial Markers for Cancer: Relevance to Diagnosis, Therapy, and Prognosis and General Understanding of Malignant Disease Mechanisms

Article

Full-text available

Nov 2012

Boel De Paepe

Cancer cells display changes that aid them to escape from cell death, sustain their proliferative powers, and shift their metabolism toward glycolytic energy production. Mitochondria are key organelles in many metabolic and biosynthetic pathways, and the adaptation of mitochondrial function has been recognized as crucial to the changes that occur in cancer cells. This paper zooms in on the pathologic evaluation of mitochondrial markers for diagnosing and staging of human cancer and determining the patients’ prognoses.

F-18-ICMT-11, a Caspase-3-Specific PET Tracer for Apoptosis: Biodistribution and Radiation Dosimetry

Article

Aug 2013

Effective anticancer therapy induces tumor cell death through apoptosis. Noninvasive monitoring of apoptosis during therapy may provide predictive outcome information and help tailor treatment. A caspase-3-specific imaging radiotracer, (18)F-(S)-1-((1-(2-fluoroethyl)-1H-[1,2,3]-triazol-4-yl)methyl)-5-(2(2,4-difluorophenoxymethyl)-pyrrolidine-1-sulfonyl)isatin ((18)F-ICMT-11), has been developed for use in PET studies. We report the safety, biodistribution, and internal radiation dosimetry profiles of (18)F-ICMT-11 in 8 healthy human volunteers. (18)F-ICMT-11 was intravenously administered as a bolus injection (mean ± SD, 159 ± 2.75 MBq; range, 154-161 MBq) to 8 healthy volunteers (4 men, 4 women). Whole-body (vertex to mid thigh) PET/CT scans were acquired at 6 time points, up to 4 h after tracer injection. Serial whole blood, plasma, and urine samples were collected for radioactivity measurement and radiotracer stability. In vivo (18)F activities were determined from quantitative analysis of the images, and time-activity curves were generated. The total numbers of disintegrations in each organ normalized to injected activity (residence times) were calculated as the area under the curve of the time-activity curve, normalized to injected activities and standard values of organ volumes. Dosimetry calculations were then performed using OLINDA/EXM 1.1. Injection of (18)F-ICMT-11 was well tolerated in all subjects, with no serious tracer-related adverse events reported. The mean effective dose averaged over both men and women was estimated to be 0.025 ± 0.004 mSv/MBq (men, 0.022 ± 0.004 mSv/MBq; women, 0.027 ± 0.004 mSv/MBq). The 5 organs receiving the highest absorbed dose (mGy/MBq), averaged over both men and women, were the gallbladder wall (0.59 ± 0.44), small intestine (0.12 ± 0.05), upper large intestinal wall (0.08 ± 0.07), urinary bladder wall (0.08 ± 0.02), and liver (0.07 ± 0.01). Elimination was both renal and via the hepatobiliary system. (18)F-ICMT-11 is a safe PET tracer with a dosimetry profile comparable to other common (18)F PET tracers. These data support the further development of (18)F-ICMT-11 for clinical imaging of apoptosis.

On the brotherhood of mitochondrial chaperones, mortalin/mtHsp70 and Hsp60

Article

Full-text available

Jan 2005

The heat shock chaperones mortalin/mitochondrial heat shock protein 70 (mtHsp70) and Hsp60 are found in multiple subcellular sites and function in the folding and intracellular trafficking of many proteins. The chaperoning activity of these 2 proteins involves different structural and functional mechanisms. In spite of providing an excellent model for an evolutionarily conserved molecular ''brotherhood,'' their individual functions, although overlapping , are nonredundant. As they travel to various locations, both chaperones acquire different binding partners and exert a more divergent involvement in tumorigenesis, cellular senescence, and immunology. An understanding of their functional biology may lead to novel designing and development of therapeutic strategies for cancer and aging.

The Multiple Roles and Therapeutic Potential of HSP60 in Cancer

Article

May 2022
BIOCHEM PHARMACOL

The molecular chaperone protein HSP60 is mainly distributed in mitochondria and assists protein folding under physiological and pathological conditions. Accumulating evidence suggests abnormally expressed HSP60 in cancer is associated with clinicopathological features and prognosis of cancer patients. HSP60 could be used as a new biomarker for both diagnostic and prognostic purpose and tumor therapy. In this review article, we briefly described the structure, functional cycle, and regulatory mechanism of HSP60, and summarized its functional diversity in cancer as well as recent progress related to the diagnostic application of HSP60 and inhibitors against HSP60, which could provide us a comprehensive understanding about the value of HSP60 in tumor management.

Chaperonin Hsp60 and Cancer Therapies

Chapter

Full-text available

Jul 2020

Introduction The heat shock protein 60 (Hsp60) is a chaperonin belonging to the chaperoning (chaperone) system that typically contributes to protein homeostasis inside mitochondria, but also plays various non-canonical roles unrelated to protein quality control beyond the organelle. Chaperonopathies are disorders in which chaperones play an etiologic-pathogenic role and contribute to the onset/progression of disease. Hsp60 chaperonopathies by mistake are diseases in which the chaperonin is apparently normal (as far as it can be determined with current methodologies) but it actively contributes to pathology, for example in certain types of cancer, and autoimmune and chronic inflammatory disorders. In certain cancers, Hsp60 is associated with the onset of malignancy and metastasization, although the mechanisms are poorly understood. In this chapter, we summarize findings on Hsp60 quantitative changes and distribution alterations in cells and tissues accompanying tumor initiation and progression. We also discuss the potential of HSP60-based anti-cancer therapies that are currently being investigated.

Cardiac Myopathy in Conditional Hsp60 Transgenic Mice

Chapter

Oct 2019

The mitochondrial chaperonin Hsp60 (Hspd1) plays important roles in sustaining cellular viability, regulate cellular functions and maintain homeostasis. Mutations in the Hsp60 gene or erratic expression has been frequently observed in wide-ranging human diseases. Targeting Hsp60 to ameliorate the prognosis of mitochondrial dysfunction-related diseases were proposed in the past. Genetically engineered mice provide a compelling tool to investigate the aetiology and pathogenesis of these diseases. Eventually, this will benefit the development of therapeutics towards these physiological complications. Conventional Hsp60 transgenic mice are often neonatally lethal. We’ve generated a unique conditional Hsp60 transgenic (Tg) mouse model to investigate the mitochondrial activities and demonstrated that ubiquitous expression of human Hsp60 protein in mice leads to neonatal death due to septum defect (ASD) and cardiac myopathy. This chapter concisely reviews recent advances regarding manipulating Hsp60 levels in cells and mouse models along with depicts our quest to develop transgenic mice to study Hsp60-related human diseases.

Evaluation of Heat Shock Protein 60 (HSP60) Chaperonin in Oncology

Chapter

Oct 2019

İsmail Ağababaoğlu

Heat shock proteins (Hsp) are a group of chaperonin that are increased production at cellular level in cellular stress situations. These stress include such as heat, infection, inflammation, many toxins such as ethanol, arsenic, some metals, ultraviolet light, some oncogenes. Under physiological conditions, Hsp helps the newly produced proteins in the cell in folding correctly. They provide stabilization of the mitochondria as a mechanism that prevents the cell from apoptosis. They function by interacting with proteins called chaperonin in the cell cycle and stabilization of mitochondria. And it protects the cell from apoptosis and directs it to the process of carcinogenesis. Therefore, it has great potential in cancer studies in many stages. Thus, our chapter aims to briefly evaluate up to date knowledge for Hsp in oncological field.

Hsp60 in Cancer Immunity: Biological Basis, Diagnostic Potential and Therapeutic Opportunities

Chapter

Oct 2019

Christian Gomez

Hsp60 is involved in tumor immune mechanisms leading to recognition of transformed cells and inhibition of growth of neoplastic tissue. As tumors grow, Hsp60 participates in disease progression through its involvement in immunoescape. In this chapter, the interactions between Hsp60 and the tumor microenvironment are discussed as they offer key elements to illustrate the context-dependent functions of Hsp60 in tumor immunomodulation. Next, the applicability of Hsp60 as a diagnostic, prognostic, and marker for therapy response is discussed. Finally, the prospect of targeting Hsp60 and its immunomodulatory effects in tumors is explored. A critical component of self- and nonself-recognition, Hsp60 has value as a tumor marker and therapeutic agent. Study of its context-dependent immune functions offer the prospect of delivering better diagnostic and personalized therapeutic approaches.

S5 Table

Data

Full-text available

Apr 2018

Biological relevance of selected cognate DFTD antigens identified by mass spectrometry analyses. (PDF)

Prognostic role of HSPs in human gastrointestinal cancer: A systematic review and meta-analysis

Article

Full-text available

Jan 2018

Background Heat shock proteins (HSPs) have been reported to be overexpressed in a wide range of human tumors. It has been shown that HSPs act as an oncogenic regulator and are involved in tumorigenesis. The clinical and prognostic significance of HSPs in gastrointestinal cancers (GICs) remains controversial. The aim of this study was to conduct a meta-analysis to assess the prognostic value of HSPs in GICs. Materials and methods A literature search was performed in PubMed, Cochrane Library, Web of Science, and Embase databases. Data on the relationship between expression of HSPs and survival outcomes were extracted. Pooled hazard ratios (HRs) with 95% CI were calculated. Results The expression of HSPs was not associated with the overall survival (OS) of GIC patients; however, it was significantly associated with worse OS for gastric cancer (GC) and colorectal cancer (CRC) patients. Conclusion Current evidence suggests that a high level of HSPs may not be a potential marker to predict the survival rate for every type of GICs. However, the expression of HSPs may predict a poor prognosis for GC and CRC patients.

Chaperonotherapy for Alzheimer’s Disease: Focusing on HSP60

Chapter

Jan 2015

This review will analyze growing evidence suggesting a convergence between two major areas of research: Alzheimer’s disease (AD) and chaperonopathies. While AD is a widely recognized medical, public health, and social problem, the chaperonopathies have not yet been acknowledged as a related burden of similar magnitude. However, recent evidence collectively indicates that such possibility exists in that AD, or at least some forms of it, may indeed be a chaperonopathy. The importance of considering this possibility cannot be overemphasized since it provides a novel point of view to examine AD and potentially suggests new therapeutic avenues. In this review, we focus on the mitochondrial chaperone HSP60 and discuss some of its biological, molecular, and pathological facets as they pertain to AD. We further illustrate how HSP60 may be an etiologic-pathogenic factor in AD and, as such, it could become a novel, effective therapeutic target. This possibility is discussed both in the light of negative chaperonotherapy, namely the development of means to inhibit HSP60 in the event its excessive activity is a disease-promoting event in AD, as well as positive chaperonotherapy, that is boosting its activity if, on the other hand, it is demonstrated that HSP60 insufficiency is a key feature of AD with such pathological consequences as causing mitochondrial dysfunction.

The tumor promoting roles of HSP60 and HIF2α in gastric cancer cells

Article

Jan 2016

The roles of HSP60 and HIF2α in diagnosis, prognosis, and prevention and treatment of various human cancers have been detected. However, the combined roles of HSP60 and HIF2α on the prognosis of patients with gastric cancer remain unclear. In this work, we confirmed that the levels of HSP60 and HIF2α messenger RNA (mRNA) and protein were higher in gastric cancer tissues than that in matched normal tissues by using real-time PCR and Western blot. Furthermore, we confirmed that inhibition of HSP60 or HIF2α could induce apoptosis and inhibit cell mobility. Co-immunoprecipitation (co-IP) was performed to determine the interaction between HSP60 and HIF2α. Lastly, we confirmed that knockdown of HSP60 or HIF2α induced apoptosis in gastric cancer cells is negatively related to the MEK/ERK signaling in vitro. In summary, HSP60 or HIF2α protein expression may be a predictive marker for the prognosis of the patients with gastric cancer. Targeting HSP60 and HIF2α could be a future strategy to improve survival of gastric patients with poor prognosis.

The Prognostic Impact of Heat Shock Proteins Expression in Patients with Esophageal Cancer: A Meta-Analysis

Article

Full-text available

Nov 2015

Purpose Heat shock proteins (HSPs) are highly conserved molecular chaperones. There are various studies that assess the prognostic value of HSPs in patients with esophageal cancer, but the conclusion remains controversial. This is the first meta-analysis study aiming to summarize the evidence on the suitability of HSPs to predict patients' survival. Materials and Methods Searching PubMed, Web of science and Medline until May 31, 2014, data were compared for overall survival in patients with down-regulated HSPs level with those with up-regulated level. We conducted a meta-analysis of 9 studies (801 patients) that correlated HSPs levels with overall survival. Data were synthesized with hazard ratios (HRs). Results The estimated risk of death was 2.93-fold greater in HSP27 negative patients than HSP27 positive patients [95% confidence interval (CI), 1.12-7.62]. When limited to esophageal squamous cell carcinoma (ESCC), the risk of death in HSP27 negative patients seemed more significant (HR, 3.90; 95% CI, 2.35-6.49). Decreased expression of HSP70 was also associated with worse survival in esophageal cancer (HR, 2.83; 95% CI, 1.90-4.23) and, when limited to ESCC, HR was 3.21 (95% CI, 1.94-5.30). Data collected, however, were not sufficient to determine the prognostic value of HSP90 in patients with ESCC nor esophageal adenocarcinomas (EADC). Conclusion In this meta-analysis, reduced HSP27 and HSP70 expressions were associated with poor survival in patients with esophageal cancer, especially esophageal squamous cell carcinoma.

Protein profile of human hepatocarcinoma cell line SMMC-7721: Identification and functional analysis

Article

May 2007
WORLD J GASTROENTERO

Yi Feng

AIM: To investigate the protein profile of human hepatocarcinoma cell line SMMC-7721, to analyze the specific functions of abundant expressed proteins in the processes of hepatocarcinoma genesis, growth and metastasis, to identify the hepatocarcinoma-specific biomarkers for the early prediction in diagnosis, and to explore the new drug targets for liver cancer therapy. METHODS: Total proteins from human hepatocarcinoma cell line SMMC-7721 were separated by two-dimensional electrophoresis (2DE). The silver-stained gel was analyzed by 2DE software Image Master 2D Elite. Interesting protein spots were identified by peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: We obtained protein profile of human hepatocarcinoma cell line SMMC-7721. Among the twenty-one successfully identified proteins, mitofilin, endoplasmic reticulum protein ERp29, ubiquinol-cytochrome C reductase complex core protein I, peroxisomal enoyl CoA hydratase, peroxiredoxin-4 and probable 3-oxoacid CoA transferase 1 precursor were the six novel proteins identified in human hepatocarcinoma cells or tissues. Specific functions of the identified heat-shock proteins were analyzed in detail, and the results suggested that these proteins might promote tumorigenesis via inhibiting cell death induced by several cancer-related stresses or via inhibiting apoptosis at multiple points in the apoptotic signal pathway. Other identified chaperones and cancer-related proteins were also analyzed. CONCLUSION: Based on the protein profile of SMMC-7721 cells, functional analysis suggests that the identified chaperones and cancer-related proteins have their own pathways to contribute to the tumorigenesis, tumor growth and metastasis of liver cancer. Furthermore, proteomic analysis is indicated to be feasible in the cancer study.

Mitochondrial Lon regulates apoptosis through the association with Hsp60–mtHsp70 complex

Article

Full-text available

Feb 2015

Human Lon protease is a mitochondrial matrix protein with several functions including protein degradation, mitochondrial DNA (mtDNA) binding, and chaperone activity. Lon is currently emerging as an important regulator of mitochondria-contributed tumorigenesis due to its overexpression in cancer cells. To understand the mechanism of increased Lon in tumor cells, we studied the interactome to identify the chaperone Lon-associated proteins by proteomics approaches using the cells overexpressing Lon. In the present study, we designed a method connecting co-immunoprecipitation (Co-IP) to in-solution digestion for the shotgun mass spectrometry. We identified 76 proteins that were putative Lon-associated proteins participated in mitochondrial chaperone system, cellular metabolism and energy, cell death and survival, and mtDNA stability. The association between Lon and NDUFS8 or Hsp60-mtHsp70 complex was confirmed by Co-IP and immunofluorescence co-localization assay. We then found that the protein stability/level of Hsp60-mtHsp70 complex depends on the level of Lon under oxidative stress. Most importantly, the ability of increased Lon-inhibited apoptosis is dependent on Hsp60 that binds p53 to inhibit apoptosis. These results suggest that the mechanism underlying cell survival regulated by Lon is mediated by the maintenance of the protein stability of Hsp60-mtHsp70 complex. This new knowledge of chaperone Lon interactome will allow us to better understand the cellular mechanism of Lon in mitochondrial function and of its overexpression in enhancing cell survival and tumorigenesis.

Inhibition of heat shock protein 90 suppresses squamous carcinogenic progression in a mouse model of esophageal cancer

Article

Jan 2015

Purpose: Heat shock protein 90 (Hsp90), a potential therapeutic target, has been widely recognized in vitro and in vivo in immunodeficient mice. Here, we aimed to evaluate the role of Hsp90 in an immunocompetent mouse model of esophageal squamous cell cancer (ESCC). Methods: The carcinogen 4-nitroquinoline 1-oxide (4NQO) was used to induce ESCC in C57BL/6 mice. Cancer progression was analyzed through observation of appearance, hematoxylin-eosin staining, immunohistochemical detection, and terminal dUTP nick-end labeling analysis. Results: 4NQO led to the progressive appearance of preneoplastic and tumoral lesions in the esophagus, with 100 % incidence of ESCC in situ occurring only after 16 weeks of carcinogen exposure. Most of these lesions evolved spontaneously into highly invasive ESCC even after 4NQO withdrawal (weeks 16-22). Interestingly, there was marked upregulation of Hsp90 and its client proteins in tumoral lesions at 22 weeks. Hsp90 inhibition by intraperitoneal injection of SNX-2112 over the following 2 weeks downregulated AKT and cyclin D1 expression, leading to significant reduction in tumor incidence and prevention of ESCC progression. Moreover, SNX-2112 treatment decreased proliferating cell nuclear antigen expression and increased the number of apoptotic cells in ESCC tissues. Conclusions: Our in vivo findings support the contribution of Hsp90 to ESCC progression, which was achieved by stimulating apoptosis and inhibition of cell proliferation, and provide a strong rationale for further evaluation of Hsp90 inhibitors for treating ESCC.

Protein papaer

Data

Full-text available

Dec 2013

Differential Protein Expression of Lymph Node Metastases of Papillary Thyroid Carcinoma Harboring the BRAF Mutation

Article

Oct 2013
ANTICANCER RES

The prognostic role of the T1799A BRAF mutation is controversial. We investigated the protein expression in papillary thyroid carcinoma (PTCs) samples harboring the specific mutation using proteomic tools. We performed two-dimensional gel electrophoresis to identify differential protein expression regarding lymph node metastasis (LNM). Proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Immunohistochemical staining was performed for 38 PTCs harboring the mutation. We validated the association between these proteins and clinicopathological factors in a test set of 121 PTCs. The expression of vimentin was increased in PTCs with LNM, but the one for HSP60, SOD2 and PEBP1 was increased in samples without LNM. HSP60 protein was up-regulated in PTCs without LNM (84.2% vs. 36.8%. p=0.003) and in PTCs without LNM harboring the mutation (58% vs. 41.8%. p=0.003) in the test set as shown by immunohistochemical staining. HSP60 protein expression may inhibit LNM in PTCs harboring the BRAF mutation and may be a useful prognostic marker.

Toll-like receptor 4 activation in Barrett’s esophagus results in a strong increase in COX-2 expression

Article

Aug 2013
J GASTROENTEROL

Barrett's esophagus (BE) is known to progress to esophageal adenocarcinoma in a setting of chronic inflammation. Toll-like receptor (TLR) 4 has been linked to inflammation-associated carcinogenesis. We aimed to determine the expression and functional activity of TLR4 in the esophagus and whether TLR4 activation in BE could promote carcinogenesis by inducing COX-2 expression. TLR4 expression in esophageal adenocarcinoma, BE, duodenum, reflux esophagitis and normal squamous esophagus biopsies was assessed using real-time PCR and validated by in situ hybridization and immunohistochemistry. Ex vivo cultures of BE, duodenum and normal squamous esophagus biopsies and a BE cell line (BAR-T) were stimulated with the TLR4 agonist lipopolysaccharide (LPS). To evaluate the effect of TLR4 activation, NF-κB activation, IL8 secretion and expression and COX-2 expression were determined. TLR4 expression was significantly increased in esophageal adenocarcinoma, BE, duodenum and reflux esophagitis compared to normal squamous esophagus. LPS stimulation resulted in NF-κB activation and a dose-dependent increase of IL8 secretion and mRNA expression. The induction of IL8 was more evident in BE compared to normal squamous esophagus. Upon LPS stimulation, COX-2 expression increased significantly in ex vivo cultured BE biopsies, which was observed in both epithelium and lamina propria cells. However, no effect was found in duodenum and normal squamous esophagus biopsies. TLR4 activation in BE results in a strong increase in COX-2 and may contribute to malignant transformation.

HSP MOLECULAR CHAPERONES IN CANCER BIOGENESIS AND ...

Data

Full-text available

Jan 2012

Giovanni Tomasello

Molecular chaperones, many of which are heat-shock proteins (HSPs), are an important class of molecules with various functions. Pathological conditions in which chaperones become etiological and/or pathogenic factors are called chaperonopathies, and are classified into by defect, by excess, and by 'mistake'. In the latter case, the chaperone is structurally and functionally normal but participates in pathways that favor disease, although in some cases the chaperone may have post-translational modifications that may lead it to change its location and function and, thus, to become pathogenic. For example, HSP-chaperones are involved in carcinogenesis in various ways, so that some forms of cancer may be considered 'chaperonopathies by mistake'. This concept suggests new strategies for anticancer therapy (chaperonotherapy), in which the primary targets or therapeutic agents are chaperones. Chaperonotherapy consists of the utilization of HSP-chaperones for treating chaperonopathies, including cancer. Negative chaperonotherapy is aimed at eliminating or blocking the action of chaperones that favor carcino-genesis or other diseases, whereas positive chaperono-therapy uses chaperones, genes or proteins, to fight against diseases, such as cancer, by stimulating the immune system or the cellular defenses against stress.

Aberrant patterns of DNA methylation, chromatin formation and gene expression in cancer

Article

Full-text available

Mar 2001

Gene function in cancer can be disrupted either through genetic alterations, which directly mutate or delete genes, or epigenetic alterations, which alter the heritable state of gene expression. The latter events are mediated by formation of transcriptionally repressive chromatin states around gene transcription start sites and an associated gain of methylation in normally unmethylated CpG islands in these regions. The genes affected include over half of the tumor suppressor genes that cause familial cancers when mutated in the germline; the selective advantage for genetic and epigenetic dysfunction in these genes is very similar. The aberrant methyla-tion can begin very early in tumor progression and mediate most of the important pathway abnormalities in cancer including loss of cell cycle control, altered function of transcription factors, altered receptor function, disruption of normal cell–cell and cell–substratum interaction, inactivation of signal transduction pathways, loss of apoptotic signals and genetic instability. The active role of the aberrant methylation in transcriptional silencing of genes is becoming increasingly understood and involves a synergy between the methylation and histone deacetylase (HDAC) activity. This synergy can be mediated directly by HDAC interaction with DNA methylating enzymes and by recruitment through complexes involving methyl-cytosine binding proteins. In the translational arena, the promoter hypermethylation changes hold great promise as DNA tumor markers and their potentially reversible state creates a target for cancer therapeutic strategies involving gene reactivation.

Prognostic significance of heat shock proteins 27 and 70 in patients with squamous cell carcinoma of the esophagus

Article

Full-text available

Apr 1999
CANCER-AM CANCER SOC

BACKGROUND Heat shock proteins (HSPs) first were defined as proteins induced by heat shock and other environmental and pathophysiologic stresses and are implicated in protein-protein interactions such as folding, translocation, and prevention of inappropriate protein aggregation. Many of their functions suggest that they play important roles in cancer.METHODS Immunohistochemical study for HSP 27 and HSP 70 was performed on buffered formalin fixed, paraffin embedded sections of 102 esophageal squamous cell carcinoma specimens using monoclonal anti-HSP 27 antibody and anti-HSP 70 antibody.RESULTSNormal squamous cells expressed both HSP 27 and HSP 70 with the exception of the basal layer. In cancerous tissue, expression of HSP 27 was evaluated as positive (+) (39 cases; 38%), reduced (±) (53 cases; 52%), or negative (−) (10 cases; 10%) and expression of HSP 70 was evaluated as (+) (14 cases; 14%), (±) (57 cases; 56%), or (−) (31 cases; 30%). There was a strong correlation between the expression of HSP 27 and HSP 70 (P < 0.0001). When compared with clinicopathologic features, expression of both HSP 27 and HSP 70 correlated negatively with lymph node metastases (P < 0.05), but not with depth of invasion or histologic grade. The reduction of the HSPs was associated significantly with poor postoperative survival (P < 0.0001). In addition, multivariate analysis revealed that HSP 27 (−) was the strongest prognostic factor among the clinicopathologic features.CONCLUSIONS This study suggests that the expression of HSP 27 and HSP 70 frequently is reduced in patients with esophageal squamous cell carcinoma and therefore should be considered an independent prognostic factor of this disease. Cancer 1999;85:1649–57. © 1999 American Cancer Society.

Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis

Article

Full-text available

May 1999

The activation of caspases represents a critical step in the pathways leading to the biochemical and morphological changes that underlie apoptosis. Multiple pathways leading to caspase activation appear to exist and vary depending on the death-inducing stimulus. We demonstrate that the activation of caspase-3, in Jurkat cells stimulated to undergo apoptosis by a Fas-independent pathway, is catalyzed by caspase-6. Caspase-6 was found to co-purify with caspase-3 as part of a multiprotein activation complex from extracts of camptothecin-treated Jurkat cells. A biochemical analysis of the protein constituents of the activation complex showed that Hsp60 was also present. Furthermore, an interaction between Hsp60 and caspase-3 could be demonstrated by co-immunoprecipitation experiments using HeLa as well as Jurkat cell extracts. Using a reconstituted in vitro system, Hsp60 was able to substantially accelerate the maturation of procaspase-3 by different upstream activator caspases and this effect was dependent on ATP hydrolysis. We propose that the ATP-dependent 'foldase' activity of Hsp60 improves the vulnerability of pro-caspase-3 to proteolytic maturation by upstream caspases and that this represents an important regulatory event in apoptotic cell death.

Distribution of an estrogen-induced protein with a molecular weight of 24,000 in normal and malignant human tissues and cells

Article

Full-text available

Apr 1983

We have shown previously that estradiol regulates the synthesis of a M(r) 24,000 protein, of unknown function, in the human breast cancer cell line MCF-7. We have also reported the preparation of monoclonal antibodies against M(r) 24,000 protein, antibodies that reacted by immunohistochemistry with MCF-7 cells and certain human breast tumor samples but not with an estrogen receptor-negative cell line. In the present paper, we report much more extensive information about cell and tissue distribution of the M(r) 24,000 protein. Several human tissue biopsy samples and human cell lines were studied by immunohistochemistry. The results show that M(r) 24,000 protein is not a fetal-associated antigen nor a milk-related protein. It was occasionally found in carcinomas other than breast tumors. It was also detected in normal estrogen target organs: uterus; oviduct; vagina; and breast. Some normal skin samples were likewise positive. The monoclonal antibodies to M(r) 24,000 protein also reacted specifically with maternal decidual cells in placenta, suggesting that these antibodies may be useful in the study of decidual transformation. The M(r) 24,000 protein was detected in six estrogen and progesterone receptor-positive cell lines but not in receptor-negative cell lines. The fraction of M(r) 24,000-positive cells in two breast cancer cell lines increased significantly after estradiol treatment. The finding of M(r) 24,000 protein in organs with secretory capacity, the varying degrees of immunostaining within the same positive cell population as expected for cells in alternating synthetic and secretory phases, and the increase of M(r) 24,000 protein immunostaining after estradiol treatment are all consistent with the possibility that M(r) 24,000 has a secretory function regulated in part by estrogens. Moreover, since decidual cells strongly reacted with the monoclonal antibodies to M(r) 24,000 protein, it is possible that the protein is under progesterone as well as estrogen control. These results point to the usefulness of these monoclonal antibodies for detecting the M(r) 24,000 protein and for studying M(r) 24,000 protein as a marker for the mechanism of hormone action in human estrogen target organs, tumors, and cell lines.

Cells in Stress: Transcriptional Activation of Heat Shock Genes

Article

Full-text available

Apr 1993

Richard I. Morimoto

In situ Apoptotic and Proliferation Index in Laryngeal Squamous Cell Carcinomas

Article

Full-text available

Feb 1998
Anal Cell Pathol

The rate of cellular growth is mainly influenced by the balance between cell proliferation and cellular decay. Since to our knowledge, no study so far has analysed the rate of proliferation and apoptosis in the normal laryngeal mucosa and in invasive laryngeal carcinomas, we performed a morphological analysis on both parameters in biopsies from 30 patients with laryngeal carcinoma. We applied the TUNEL end labelling technique for the investigation of apoptosis and immunohistochemistry (Ki‐67 antigen) for the determination of the cell proliferation. In our study we demonstrated that invasive tumour growth of the larynx coincides with an increase of both cellular proliferation and apoptosis. Both parameters, however, affected various tumour areas differently. While there was a preferential expression of the Ki‐67 antigen at the tumour–stroma interface, apoptotic figures could be found randomly distributed in the tumour. This indicates that the replication of tumour cells and tumour cell decay are differently distributed and possibly independently regulated. Since we observed a particularly strong increase of cell proliferation at the tumour–stroma interface which outnumbered the corresponding rate of apoptosis by far, the enhanced cell proliferation at the tumour border seems to be a main factor for tumour growth. A statistical evaluation revealed significant correlation between the apoptotic index and the degree of tumour cell differentiation, indicating that a high rate of apoptosis coincides with a high level of tumour cell differentiation. There was, however, no statistically significant correlation between prognostic clinical parameters and the rate of apoptosis or that of proliferation.

Presence of a pre-apoptotic complex of pro-caspase-3, Hsp60 and Hsp10 in the mitochondrial fraction of Jurkat cells

Article

Full-text available

May 1999

Activation of pro-caspase-3 is a central event in the execution phase of apoptosis and appears to serve as the convergence point of different apoptotic signaling pathways. Recently, mitochondria were found to play a central role in apoptosis through release of cytochrome c and activation of caspases. Moreover, a sub-population of pro-caspase-3 has been found to be localized to this organelle. In the present study, we demonstrate that pro-caspase-3 is present in the mitochondrial fraction of Jurkat T cells in a complex with the chaperone proteins Hsp60 and Hsp10. Induction of apoptosis with staurosporine led to the activation of mitochondrial pro-caspase-3 and its dissociation from the Hsps which were released from mitochondria. The release of Hsps occurred simultaneously with the release of other mitochondrial intermembrane space proteins including cytochrome c and adenylate kinase, prior to a loss of mitochondrial transmembrane potential. In in vitro systems, recombinant Hsp60 and Hsp10 accelerated the activation of pro-caspase-3 by cytochrome c and dATP in an ATP-dependent manner, consistent with their function as chaperones. This finding suggests that the release of mitochondrial Hsps may also accelerate caspase activation in the cytoplasm of intact cells.

Negative regulation of the APAF-1 apoptosome by HSP70

Article

Full-text available

Sep 2000

Release of cytochrome c from mitochondria by apoptotic signals induces ATP/dATP-dependent formation of the oligomeric Apaf-1-caspase-9 apoptosome. Here we show that the documented anti-apoptotic effect of the principal heat-shock protein, Hsp70, is mediated through its direct association with the caspase-recruitment domain (CARD) of Apaf-1 and through inhibition of apoptosome formation. The interaction between Hsp70 and Apaf-1 prevents oligomerization of Apaf-1 and association of Apaf-1 with procaspase-9. On the basis of these results, we propose that resistance to apoptosis exhibited by stressed cells and some tumours, which constitutively express high levels of Hsp70, may be due in part to modulation of Apaf-1 function by Hsp70.

Pandey P, Saleh A, Nakazawa A, Kumar S, Srinivasula SM, Kumar V, Weichselbaum R, Nalin C, Alnemri ES, Kufe D, Kharbanda SNegative regulation of cytochrome c-mediated oligomerization of Apaf-1 and activation of procaspase-9 by heat shock protein 90. EMBO J 19: 4310-4322

Article

Full-text available

Sep 2000

The release of cytochrome c from mitochondria results in the formation of an Apaf-1-caspase-9 apoptosome and induces the apoptotic protease cascade by activation of procaspase-3. The present studies demonstrate that heat shock protein 90 (Hsp90) forms a cytosolic complex with Apaf-1 and thereby inhibits the formation of the active complex. Immunodepletion of Hsp90 depletes Apaf-1 and thereby inhibits cytochrome c-mediated activation of caspase-9. Addition of purified Apaf-1 to Hsp90-depleted cytosolic extracts restores cytochrome c-mediated activation of procaspase-9. We also show that Hsp90 inhibits cytochrome c-mediated oligomerization of Apaf-1 and thereby activation of procaspase-9. Furthermore, treatment of cells with diverse DNA-damaging agents dissociates the Hsp90-Apaf-1 complex and relieves the inhibition of procaspase-9 activation. These findings provide the first evidence for a negative cytosolic regulator of cytochrome c-dependent apoptosis and for involvement of a chaperone in the caspase cascade.

Human Class I Histone Deacetylase Complexes Show Enhanced Catalytic Activity in the Presence of ATP and Co-immunoprecipitate with the ATP-dependent Chaperone Protein Hsp70

Article

Full-text available

Apr 2002

Antibodies to histone deacetylases (HDACs) have been used to immuno-isolate deacetylase complexes from HeLa cell extracts. Complexes shown to contain HDAC1, HDAC3, HDAC6, and HDAC1+2 as their catalytic subunits have been used in an antibody-based assay that detects deacetylation of whole histones at defined lysines. The class II deacetylase HDAC6 was inactive in this assay, but the three class I enzymes deacetylated all histone lysines tested, although with varying efficiency. In comparison to HDAC1, HDAC3 preferentially deacetylated lysines 5 and 12 of H4 and lysine 5 of H2A. H4 tails in purified mononucleosomes were refractory to deacetylation by both HDAC1 and HDAC3, unless ATP was added to the reaction mix. Surprisingly, ATP also consistently enhanced cleavage of free, non-nucleosomal histones, but not small peptides, by both enzyme complexes. We found no evidence that ATP operates by phosphorylation of components of the HDAC complex, but have shown that HDACs 1, 2, and 3 all co-immunoprecipitate with the ATP-dependent chaperone protein Hsp70. Another common ATP-dependent chaperone, Hsp90, was absent from all HDAC complexes tested, whereas Hsp60 associated with HDAC1 only. We suggest that Hsp chaperone proteins enhance the deacetylase activity of HDAC complexes by ATP-dependent manipulation of protein substrates.

Expression of 60-kD Heat Shock Protein Increases during Carcinogenesis in the Uterine Exocervix

Article

Full-text available

Dec 2002

The aim of the present study was to determine the presence and expression of the 60-kD heat shock protein (HSP60) in the dysplasia-carcinoma sequence in the uterine exocervix and to evaluate its diagnostic and prognostic significance. We performed Western blot and immunohistochemical analyses on biopsies from 40 cases, consisting of 10 normal exocervical biopsies, 10 low-grade squamous intraepithelial lesions (L-SIL), 10 high-grade squamous intraepithelial lesions (H-SIL) and 10 cancerous exocervices (G2 grade). The immunohistochemical results were quantified by computer-assisted image analysis. Western blot analysis showed that HSP60 was undetectable in normal tissues and that there was a gradual increase of protein expression from L-SIL to carcinoma. Immunostaining for HSP60 was negative in normal tissue and positive in basal and parabasal layers of L-SIL epithelium; H-SIL were markedly stained in all layers of epithelium, and carcinomas showed an even stronger positivity. The increasing expression correlated with the malignancy grade. Finally, koilocytes were mostly negative in L-SIL and positive in H-SIL. The increasing degree of expression of HSP60 from L-SIL to carcinoma and the different intraepithelial distribution between L-SIL and H-SIL could be used as a new diagnostic tool. Moreover, HSP60 could have a role in cervical carcinogenesis.

60KDa chaperonin (HSP60) is over-expressed during colorectal carcinogenesis

Article

Full-text available

Feb 2003

The aim of the present study was to evaluate the expression of the heat shock protein 60 (HSP60), a mitochondrial matrix-associated protein belonging to the chaperonin family, in colorectal adenomas and cancers, comparing them to normal colonic tissues and hyperplastic polyps. We performed both immunohistochemistry and Western blot analysis for HSP60. Immunohistochemistry resulted positive in all tubular adenomas and infiltrating adenocarcinomas. By contrast, normal tissues and hyperplastic polyps were negative. Quantitative analysis showed that tubular adenomas with different levels of dysplasia did not present statistical differences concerning HSP60 positivity. In addition, carcinomas always showed the highest expression. Western blot analysis confirmed these observations. These data suggest that HSP60 over-expression is an early event in carcinogenesis. We suspect that HSP60 plays a different role in colorectal carcinogenesis with respect to that in normal cells, which foresees its possible use as diagnostic and prognostic tools.

Aberrant patterns of DNA methylation, chromatin formation and gene expression in cancer

Article

Apr 2001
HUM MOL GENET

Stephen B. Baylin

Gene function in cancer can be disrupted either through genetic alterations, which directly mutate or delete genes, or epigenetic alterations, which alter the heritable state of gene expression. The latter events are mediated by formation of transcriptionally repressive chromatin states around gene transcription start sites and an associated gain of methylation in normally unmethylated CpG islands in these regions. The genes affected include over half of the tumor suppressor genes that cause familial cancers when mutated in the germline; the selective advantage for genetic and epigenetic dysfunction in these genes is very similar. The aberrant methylation can begin very early in tumor progression and mediate most of the important pathway abnormalities in cancer including loss of cell cycle control, altered function of transcription factors, altered receptor function, disruption of normal cell-cell and cell-substratum interaction, inactivation of signal transduction pathways, loss of apoptotic signals and genetic instability. The active role of the aberrant methylation in transcriptional silencing of genes is becoming increasingly understood and involves a synergy between the methylation and histone deacetylase (HDAC) activity. This synergy can be mediated directly by HDAC interaction with DNA methylating enzymes and by recruitment through complexes involving methyl-cytosine binding proteins. In the translational arena, the promoter hypermethylation changes hold great promise as DNA tumor markers and their potentially reversible state creates a target for cancer therapeutic strategies involving gene reactivation.

Detection of DNA Fragmentation in Human Breast Cancer Tissue by an Antibody Specific to Single-stranded DNA

Article

Jun 1998

While there have been many reports concerning the clinical significance of bcl-2 expression in human breast cancer, little is known about apoptosis in primary breast cancers. We immunohistochemically examined DNA fragmentation in 107 primary human breast cancers from Japanese women using an antibody specific to single-stranded DNA. The apoptosis index, calculated as the product of the positive cell number and the cellularity coefficient, ranged from 0 to 48. The average incidence of apoptosis was calculated as 0.1% of tumor cells. No relationships were observed among the apoptosis index, expression of bcl-2, and the histological grade of the tumors. Almost all apoptotic cells were phagocytosed by surrounding tumor cells immediately after DNA fragmentation. Apoptotic body formation was rare. The apoptotic cells seemed to be degraded within phagocytes, leaving no trace of apoptosis except the tiny shells of nuclei. The intensive phagocytic reaction might be one of the main reasons for the low incidence of apoptosis in human breast cancers.

High constitutive expression of heat shock protein 90 alpha in human acute leukemia cells

Article

Jun 1992
LEUKEMIA RES

The constitutive expression of the genes for four heat shock proteins (hsps) was studied in leukemia cell lines, cells obtained from patients with acute leukemia, and normal blood cells by means of Northern-blot analysis. Western-blot analysis with hsp90 antibody showed that the leukemia cells contained larger amounts of hsp90 than the normal peripheral mononuclear cells. The expression of the hsp90α gene was enhanced in the leukemia cell lines and the acute leukemia cells from patients as compared with the normal blood cells. In contrast, the expression of the hsp90β gene could hardly be recognized in either the acute leukemia cells or the normal blood cells. An increased expression of hsp70 gene was observed in only one patient. The expression of the hsp27 gene was enhanced in one-half the patients with common acute lymphoblastic leukemia. Thus, exclusively the hsp90α gene was expressed highly in the leukemia cells, indicating its association with cellular proliferation.

Mammalian stress response: Cell physiology, structure/function of stress proteins, and implications for medicine and disease

Article

Nov 1992

W J Welch

Ferrarini M, Heltai S, Zocchi MR, Rugarli CUnusual expression and localization of heat-shock proteins in human tumor cells. Int J Cancer 51: 613-619

Article

Jun 1992

It has been suggested that members of HSP families represent the surface target of immune responses leading to tumor rejection in mice. Here we report that tumor cells, compared with normal cells, constitutively expressed 2- to 10-fold higher levels of intracellular HSP90. Moreover, in the absence of environmental stress, 2 lines (out of 6) expressed the "inducible" HSP72, which was also detectable in fresh tumor cells. HSP72 expression was not regulated during the cell cycle, in contrast with what has been observed with normal cells. Both HSP90 and HSP72 proteins exhibited a heterogeneous pattern of intracellular distribution in most cells, HSP72 being confined mainly to the nuclear compartment. Finally, we could detect both HSP90 and, to a lesser extent, HSP72 (that are generally believed to be located intracellularly) at the surface of some tumor cell lines. We conclude that tumor cells differ from normal cells in their pattern of HSP expression; this might imply a role of HSPs in eliciting an immune response against cancer.

Clinical and biological significance of HSP89 alpha in human breast cancer

Article

Feb 1992

In order to isolate and characterize genes whose expression may be altered in breast malignancy, we screened a cDNA library with a polyclonal anti-serum against breast-cancer-metastasis membranes and isolated several immunopositive clones. One of these, AJ1, was analyzed in detail and found to be expressed at varying levels as a 3.3-kb mRNA in all of 143 breast cancers. High expression was associated with lymph-node involvement (p = 0.03). Comparison between high- and low-expressing groups showed a significant difference at 4 and 6 years for both overall (p = 0.004 and p = 0.002 respectively) and disease-free (p = 0.0001 and p = 0.04 respectively) survival, but not at 11 years. AJ1 was expressed at much lower levels in non-malignant biopsies as compared with malignant tissue (p = 0.001). Expression was observed in breast-cancer cell lines MCF-7, ZR-75-1, T47D, MDA-MB-231 and HBL 100. Partial sequence analysis of the 620 bp clone showed complete homology with human heat-shock protein 89 alpha. In addition to being heat-inducible in all the breast cell lines examined, AJ1 levels were increased by estradiol (blocked by cyclohexamide and tamoxifen), EGF, oxytocin and vasopressin in a time-dependent manner in MCF-7 cells and by estradiol, EGF, prolactin and hydrocortisone in T47D cells. In MDA-MB-231 cells, EGF caused down-regulation of AJ1 mRNA levels. The increasing evidence for the association of heat-shock proteins with steroid receptors suggests that AJ1 may play an important role in the control of estrogen-receptor transcriptional activity in breast cancers.

The Heat-Shock Response

Article

Feb 1986

S.L. Lindquist

Cell Death: The Significance of Apoptosis

Article

Feb 1980
INT REV CYTOL

The classification of cell death can be based on morphological or biochemical criteria or on the circumstances of its occurrence. Currently, irreversible structural alteration provides the only unequivocal evidence of death; biochemical indicators of cell death that are universally applicable have to be precisely defined and studies of cell function or of reproductive capacity do not necessarily differentiate between death and dormant states from which recovery may be possible. It has also proved feasible to categorize most if not all dying cells into one or the other of two discrete and distinctive patterns of morphological change, which have, generally, been found to occur under disparate but individually characteristic circumstances. One of these patterns is the swelling proceeding to rupture of plasma and organelle membranes and dissolution of organized structure—termed “coagulative necrosis.” It results from injury by agents, such as toxins and ischemia, affects cells in groups rather than singly, and evokes exudative inflammation when it develops in vivo. The other morphological pattern is characterized by condensation of the cell with maintenance of organelle integrity and the formation of surface protuberances that separate as membrane-bounded globules; in tissues, these are phagocytosed and digested by resident cells, there being no associated inflammation.

Apoptosis in the Pathogenesis and Treatment of Disease

Article

Apr 1995

Craig B. Thompson

In multicellular organisms, homeostasis is maintained through a balance between cell proliferation and cell death. Although much is known about the control of cell proliferation, less is known about the control of cell death. Physiologic cell death occurs primarily through an evolutionarily conserved form of cell suicide termed apoptosis. The decision of a cell to undergo apoptosis can be influenced by a wide variety of regulatory stimuli. Recent evidence suggests that alterations in cell survival contribute to the pathogenesis of a number of human diseases, including cancer, viral infections, autoimmune diseases, neurodegenerative disorders, and AIDS (acquired immunodeficiency syndrome). Treatments designed to specifically alter the apoptotic threshold may have the potential to change the natural progression of some of these diseases.

Blot WJOEsophageal cancer trends and risk factors. Semin Oncol 21: 403-410

Article

Sep 1994
SEMIN ONCOL

William J Blot

Esophageal cancer displays unique epidemiologic features that distinguish it from all other malignancies. It shows marked geographic variation both internationally, with exceptionally high rates (some of the world's highest for any cancer) in limited areas of Asia, and nationally, with clustering of increased rates within the United States as well. However, the patterns are changing with rates of squamous cell carcinomas decreasing and adenocarcinomas increasing rapidly in several western countries. The causes of the clusters of squamous cell carcinomas in parts of Asia and Africa are not well known, but within the United States and other western countries, tobacco and alcohol consumption are the major determinants. Nutritional factors also may play a major role, with diets high in fresh fruits and vegetables consistently associated with reduced risks. The causes of the rapidly increasing rates of adenocarcinomas of the esophagus, and reasons or its occurrence primarily among white men, are enigmatic. Additional research on the etiology of this emerging cell type is warranted, and may provide information crucial to the development of readily implementable preventive strategies.

Principles of Chaperone-Assisted Protein Folding: Differences Between in Vitro and in Vivo Mechanisms

Article

Jul 1996

Molecular chaperones in the eukaryotic cytosol were shown to interact differently with chemically denatured proteins and their newly translated counterparts. During refolding from denaturant, actin partitioned freely between 70-kilodalton heat shock protein, the bulk cytosol, and the chaperonin TCP1-ring complex. In contrast, during cell-free translation, the chaperones were recruited to the elongating polypeptide and protected it from exposure to the bulk cytosol during folding. Posttranslational cycling between chaperone-bound and free states was observed with subunits of oligomeric proteins and with aberrant polypeptides; this cycling allowed the subunits to assemble and the aberrant polypeptides to be degraded. Thus, folding, oligomerization, and degradation are linked hierarchically to ensure the correct fate of newly synthesized polypeptides.

The stress response and the lung

Article

Aug 1997
Am J Physiol

The stress response is a highly conserved cellular defense mechanism defined by the rapid and specific expression of stress proteins, with concomitant transient inhibition of nonstress protein gene expression. The stress proteins mediate cellular and tissue protection against diverse cytotoxic stimuli. Among the many classes of stress proteins, heat shock protein 70 and heme oxygenase-1 are the best characterized with respect to lung biology. A potential role for stress proteins in human lung disease is inferred from studies demonstrating stress protein expression in the lungs of patients with cancer, asthma, and acute lung injury. Several examples of stress protein-mediated cytoprotection exist in cell and animal models of acute lung injury. Stress protein induction protects rats against acute lung injury caused by either systemic administration of endotoxin or intratracheal administration of phospholipase A1. In vitro, increased expression of stress proteins protects lung cells against endotoxin-mediated apoptosis and oxidant injury. The mechanisms of stress response-mediated cytoprotection may involve the enzymatic and molecular chaperone properties of stress proteins. Alternatively, the stress response may protect by modulating lung proinflammatory responses. Data from extrapulmonary systems suggest that stress response-associated factors (heat shock protein 70 and heat shock factor) are directly involved in modulation of proinflammatory gene expression. Recent evidence also demonstrates interactions between the stress response and the I-kappa B/nuclear factor-kappa B pathway in cultured lung cells. Increased understanding about the role of stress proteins in lung biology may support efforts to selectively increase expression of one or more stress proteins to provide protection against human acute lung injury.

Apoptotic Index in Ovarian Carcinoma: Correlation with Clinicopathologic Factors and Prognosis

Article

Oct 1997

The apoptotic index (apoptotic cells/1000 tumor cells, AI) was evaluated in 71 ovarian carcinomas, all surgically resected. Apoptosis was examined by modified terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) method in histologic sections. High AI (>/=2.8) significantly correlated with high mitotic index (P = 0.05), high histologic grade of the tumor (P = 0. 018), and short overall survival (P = 0.017). An inverse relationship between AI and bcl-2 protein expression was also observed (P = 0.007). In addition, AI was assessed in 5 ovarian epithelial tumors of borderline malignancy, and all were categorized as low AI (<2.8). No significant correlation was found between AI and other clinicopathologic factors, such as age, clinical stage, lymph node metastasis, tumor size, histology of the tumor, and expression of p53 protein. Multivariate survival analysis showed that only clinical stage (P = 0.0395) and mitotic index (P = 0.0387) had independent prognostic value, whereas AI did not. Our results suggest that counting apoptosis can be useful for predicting the patient survival in ovarian carcinoma, although AI is not an independent prognostic factor. It is also suggested that bcl-2 protein is an important regulator of apoptosis in ovarian carcinoma.

Single-stranded DNA as an immunocytochemical marker for apoptotic change of ischemia in gerbil hippocampus

Article

Feb 1998
NEUROSCI LETT

The light and electron microscopic localizations of single stranded DNA (SSD) protein, a marker of apoptosis and programmed cell death, in the gerbil hippocampus were examined by immunocytochemistry after transient brain ischemia. SSD-immunoreactive (IR) cells appeared from post-operative day 1 (PO 1) to PO 7 after 5- or 10-min ischemia. Immunoreaction was recognized in the nucleus of the CA1 pyramidal neurons without remarkable morphological changes on PO 1. These findings suggest that SSD degradation can occur during delayed neuronal death in the CA1, preceding the appearance of double strand breaks, one of the characteristic features of apoptosis.

Apoptosis as a Prognostic Factor in Colorectal Carcinoma

Article

Feb 1998

We investigated the occurrence of apoptotic cell death in 104 colorectal carcinomas by terminal-deoxynucleotidyl-transferase-mediated dUTP-diotin nick end-labeling (TUNEL) to determine whether apoptosis could be a useful prognostic factor. The apoptotic index (AI) was calculated as the percentage of positive cancer cells per 1,000 cancer cells, the median AI being 4.1, with a range of 1.9-4.7. Apoptosis was less frequently observed in tumors with higher malignant potential, such as those at advanced stages Dukes B, C and D, than those at Dukes stage A (P < 0.05); in tumors showing evidence of moderate differentiation than in well-differentiated tumors (P < 0.05); and in tumors with venous invasion or lymph node metastasis than in those without these features (P < 0.05). Moreover, the subgroup of patients with a low AI of < 4.1 had a significantly poorer survival rate than the subgroup with a high AI in tumors at Dukes stage C, the 5-year survival rates being 33% vs 68% (P < 0.05; Cox-Mantel). Our findings suggest that less apoptosis might result in a greater progression of colorectal carcinoma, and that the rate of apoptosis might be an indicator of the degree of malignancy. Thus it would appear that the frequency of apoptosis in tumor cells could be a useful prognostic factor in colorectal carcinoma.

Abundance of heat shock proteins (HSP89, HSP60, and HSP27) in malignant cells of Hodgkin's disease

Article

Jan 1999

Heat shock proteins (HSPs) or stress proteins are synthesized by cells in response to environmental stress. Expression of HSPs by cells may have important physiological or pathological implications. In this study, we carried out an immunohistochemical and biochemical examination of low (hsp27), intermediate (hsp60), and high (hsp89) molecular weight HSP expression in reactive lymph nodes and in lymph nodes of patients with various types of lymphomas. In normal or reactive lymphoid tissues, hsp89 is abundant in large "transformed" lymphoid cells and immunoblasts. Hsp60 is widely distributed in lymphoid tissues, whereas hsp27 is absent in all lymphoid cells and histiocytes. Among lymphomas, the Hodgkin's Reed-Sternberg (H-RS) cells in Hodgkin's disease (HD) had the greatest abundance of hsp89 and hsp60 and, in 20% of cases, hsp27, in contrast to a much weaker staining of anti-hsp89 and -hsp60 in the background reactive lymphoid cells. The large lymphoid cells in small lymphocytic lymphoma are also rich in hsp89, but not hsp60 and hsp27. In contrast, the malignant cells in anaplastic large cell lymphoma and most high-grade tumors, including immunoblastic lymphomas, expressed minimal amounts of hsp89 and hsp60 and virtually no hsp27. Thus, the cellular level of HSPs was neither correlated with the proliferative capacity nor with the aggressiveness of the lymphomas. Hsp89, hsp60, and hsp27, as well, serve critical roles in the chaperoning of cellular proteins (e.g., a Mr 43,000 protein) in H-RS cells. The known interactions of HSPs with Rb, p53, peptide-MHC class II complexes, and cofactors of the glucocorticoid hormone receptor have further broadened the importance of HSPs in cell metabolism and in response to extracellular signals for proliferation, differentiation, or growth suppression (or apoptosis) of H-RS cells. Abundant HSP expression is seen only in HD, but not in other lymphomas. Such expression could have vital roles in the pathogenesis of HD.

Detection of Apoptotic Cells in Human Colorectal Cancer by Two Different in situ Methods: Antibody against Single-stranded DNA and Terminal Deoxynucleotidyl Transferase-mediated dUTP-biotin Nick End-labeling (TUNEL) Methods

Article

Mar 1999
Jpn J Canc Res

We comparatively investigated the extent of apoptotic cell loss in human colorectal cancers evaluated by two methods, namely the conventional terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end-labeling (TUNEL) method and immunohistochemistry for single-stranded (ss) DNA. The apoptotic index (AI) obtained with the TUNEL method was higher than that shown by the immunohistochemistry for ssDNA. However, a significant correlation in AIs evaluated by these methods was found. The AIs obtained by both methods were significantly higher in the advanced cancers than in the early cancers. Cellular proliferation activity was assessed in terms of positivity rate (PR) for expression of proliferating cell nuclear antigen (PCNA). The PR of advanced cancers was significantly higher than that of early cancers. The present results indicate that immunohistochemistry for ssDNA is useful (as is the TUNEL method) for evaluation of apoptotic tumor cells in colorectal carcinomas. In addition, it was confirmed that there is a remarkable increase of not only proliferation activity, but also tumor cell apoptosis in the process of progression of colon cancer from early to advanced stages of the disease.

Immunohistochemical detection of HSP60-expression in human ovarian cancer. Correlation with survival in a series of 247 patients

Article

May 1999

In a previous pilot study, HSP60 expression at the transcriptional (mRNA) level was shown to be a negative prognostic factor in ovarian cancer. The aim of this study was to determine HSP60-expression by means of immunohistochemistry and to correlate the results with survival in a large series of ovarian carcinoma patients with a closed follow-up. Slides from routinely processed, paraffin-embedded tumor blocks belonging to 247 patients with epithelial ovarian carcinoma were studied for the overexpression of HSP60 using the Lk2 monoclonal antibody and the strepatvidin-biotin-peroxidase technique. HSP60-expression was correlated with overall survival by means of life-table analysis and the Kaplan-Meier method. 47 tumors (19%) expressed HSP60. Of them, 12/29 (41.4% positivity) were stage I tumors, whereas only 35 out of the remaining 218 tumors in more advanced surgical stage (16.1%) showed HSP60 staining. This difference was statistically significant (Fisher s exact test; p = 0.004). Even when stratifying stage for stage, the difference between groups still remained statistically significant (Chi square test; p = 0.0095). The survival curve analysis showed a significant difference in favor of those tumors expressing HSP60 (median survival 28 vs. 37 months; log-rank test, p = 0.02). Immunohistochemical detection of HSP60-expression in human epithelial ovarian carcinoma is significantly more frequent in tumors from patients with initial stages of the disease. Therefore, HSP60-expression determined by this method is associated with a significantly better prognosis.

Overexpression and Localization of Heat Shock Proteins mRNA in Pancreatic Carcinoma

Article

Jul 2000

In the present study we examined the localization and overexpression of heat shock proteins (hsps), mainly hsp90, in pancreatic carcinoma tissue compared with control tissue (including chronic pancreatitis and normal pancreas tissue), with the aid of immunohistochemical staining, in situ hybridization and reverse transcriptase polymerase chain reaction. Hsp90 alpha mRNA was overexpressed more highly in pancreatic carcinoma than in the control tissue. The proliferating-cell-nuclear-antigen labeling index was also high in pancreatic carcinoma tissue compared with the other tissue. These findings suggest that the overexpression of hsp90 alpha mRNA in carcinomas may be correlated with cell proliferation. However, hsp90 beta was constitutively overexpressed almost equally in all groups of pancreatic tissue including pancreatic carcinoma, chronic pancreatitis and normal pancreas tissue. Immunohistochemical staining demonstrated a differentiation in the expression of hsp90 between histological types of pancreatic carcinoma. These findings suggest that hsp90 alpha is involved in carcinogenesis and that hsp90 beta is correlated to structural conformation. Hsp90 alpha and hsp90 beta seem to perform different functions in tissue containing malignant cells. P53, MDM2 and WAF1, that were cell-cycle-related oncogene product were more strongly expressed in the nuclei of the cancer cells of the cancer tissue. Especially, MDM2 was more strongly expressed in mucinous carcinoma and the mucin secreting tissues surrounding pancreatic carcinoma tissue. The expression of MDM2 protein might also be correlated to secretion systems during structural conformation and be correlated to hsp90 beta.

Detection of DNA Fragmentation in Human Breast Cancer Tissue by an Antibody Specific to Single-stranded DNA.

Article

Jul 1998
Breast Canc

While there have been many reports concerning the clinical significance of bcl-2 expression in human breast cancer, little is known about apoptosis in primary breast cancers. We immunohistochemically examined DNA fragmentation in 107 primary human breast cancers from Japanese women using an antibody specific to single-stranded DNA. The apoptosis index calculated as the product of the positive cell number and the cellularity coefficient, ranged from 0 to 48. The average incidence of apoptosis was calculated as 0.1% of tumor cells. No relationships were observed among the apoptosis index, expression of bcl-2, and the histological grade of the tumors. Almost all apoptotic cells were phagocytosed by surrounding tumor cells immediately after DNA fragmentation. Apoptotic body formation was rare. The apoptotic cells seemed to be degraded within phagocytes, leaving no trace of apoptosis except the tiny shells of nuclei. The intensive phagocytic reaction might be one of the main reasons for the low incidence of apoptosis in human breast cancers.

Combined and Individual Mitochondrial HSP60 and HSP10 Expression in Cardiac Myocytes Protects Mitochondrial Function and Prevents Apoptotic Cell Deaths Induced by Simulated Ischemia-Reoxygenation

Article

May 2001
CIRCULATION

The mitochondrial heat-shock proteins HSP60 and HSP10 form a mitochondrial chaperonin complex, and previous studies have shown that their increased expression exerts a protective effect against ischemic injury when cardiac myocytes are submitted to simulated ischemia. The more detailed mechanisms by which such a protective effect occurs are currently unclear. We wanted to determine whether HSP60 and HSP10 could exert a protection against simulated ischemia and reoxygenation (SI/RO)-induced apoptotic cell death and whether such protection results from decreased mitochondrial cytochrome c release and caspase-3 activation and from the preservation of ATP levels by preservation of the electron transport chain complexes. In addition, we explored whether increased expression of HSP60 or HSP10 by itself exerts a protective effect. We overexpressed HSP60 and HSP10 together or separately in rat neonatal cardiac myocytes using an adenoviral vector and then subjected the myocytes to SI/RO. Cell death and apoptosis in myocytes were quantified by parameters such as enzyme release, DNA fragmentation, and caspase-3 activation. Overexpression of the combination of HSP60 and HSP10 and of HSP60 or HSP10 individually protected myocytes against apoptosis. This protection is accompanied by decreases in mitochondrial cytochrome c release and in caspase-3 activity and increases in ATP recovery and activities of complex III and IV in mitochondria after SI/RO. These results suggest that mitochondrial chaperonins HSP60 and HSP10 in combination or individually play an important role in maintaining mitochondrial integrity and capacity for ATP generation, which are the crucial factors in determining survival of cardiac myocytes undergoing ischemia/reperfusion injury.

Expression of p53 protein related to smoking and alcoholic beverage drinking habits in patients with esophageal cancers

Article

Jun 2001
CANCER LETT

In esophageal squamous cell carcinoma (SCC), we used immunohistochemical analysis to further elucidate the correlation of p53 protein expression with clinicopathological factors, as well as with risk factors, such as tobacco smoking, alcohol consumption and a family history of cancer, using odds ratios (ORs). The expression of p53 protein was demonstrated in 55.1% of 89 esophageal SCC cases examined by immunohistochemistry. The expression of p53 protein did not correlate with gender, age, histological grading, lymph node metastasis, or TNM stage. The prevalence of p53 expression was significantly higher in patients with multiple primary esophageal cancers (P<0.05). p53 expression did not correlate with prognosis in univariate survival analysis. The esophageal SCC in either smokers or alcohol users was 4.67-5.83 times more likely to express p53 protein, while the likelihood of p53 expression in patients who use both tobacco and alcohol was more than 14.0 times. However, a significant association was not found between p53 expression and a family history of cancer, this having an OR as low as 1.85. The expression of p53 protein did not correlate with clinicopathological factors and prognosis in univariate and multivariate survival analyses. In contrast, tobacco smoking and alcohol consumption were shown to be strongly associated with p53 mutations in esophageal carcinogenesis.

Apoptosis as an independent prognostic indicator in squamous cell carcinoma of the esophagus

Article

Aug 2001

Apoptosis plays a crucial role in determining net cell proliferation and cell turnover in various tumors. The rate of apoptosis in tumor cells has been reported to be a useful prognostic indicator in colorectal carcinoma. We examined apoptosis in 72 specimens of esophageal squamous cell carcinoma, by the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) digoxigenin-nick end labeling (TUNEL) method. We examined correlation of apoptosis with outcome, clinicopathological features, and expression of the apoptosis-related proteins p53 and Bcl-2. The percentage of apoptotic cells, or apoptotic index (AI), ranged from 0.8 to 9.4 (mean: 3.47; SD: 2.02). Overall, 5-year survival of patients with high AI (AI > or = 5.0; n = 18) tumors was significantly higher than that of patients with low AI tumors (AI < 5.0; n = 58; 76.9% versus 44.9%; P = 0.042). AI did not correlate significantly with the clinicopathological features of patient age and sex, depth of tumor and histological differentiation, lymph node metastasis, lymphatic invasion, or venous invasion. In p53-negative tumors, the AI was significantly higher than in p53-positive tumors. We concluded that AI may be a useful prognostic indicator in esophageal squamous cell carcinoma following curative surgery, and that apoptosis in this tumor is related to relative underexpression of p53 protein.

Jiang J, Ballinger CA, Wu Y, Dai Q, Cyr DM, Hohfeld J and Patterson CCHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J. Biol. Chem. 276: 42938-42944

Article

Dec 2001

Proper folding of proteins (either newly synthesized or damaged in response to a stressful event) occurs in a highly regulated fashion. Cytosolic chaperones such as Hsc/Hsp70 are assisted by cofactors that modulate the folding machinery in a positive or negative manner. CHIP (carboxyl terminus of Hsc70-interacting protein) is such a cofactor that interacts with Hsc70 and, in general, attenuates its most well characterized functions. In addition, CHIP accelerates ubiquitin-dependent degradation of chaperone substrates. Using an in vitro ubiquitylation assay with recombinant proteins, we demonstrate that CHIP possesses intrinsic E3 ubiquitin ligase activity and promotes ubiquitylation. This activity is dependent on the carboxyl-terminal U-box. CHIP interacts functionally and physically with the stress-responsive ubiquitin-conjugating enzyme family UBCH5. Surprisingly, a major target of the ubiquitin ligase activity of CHIP is Hsc70 itself. CHIP ubiquitylates Hsc70, primarily with short, noncanonical multiubiquitin chains but has no appreciable effect on steady-state levels or half-life of this protein. This effect may have heretofore unanticipated consequences with regard to the chaperoning activities of Hsc70 or its ability to deliver substrates to the proteasome. These studies demonstrate that CHIP is a bona fide ubiquitin ligase and indicate that U-box-containing proteins may comprise a new family of E3s.

Expression of heat shock protein 90 beta in human gastric cancer tissue and SGC7901/VCR of MDR-type gastric cancer cell line

Article

Dec 1999

To examine the expression of heat shock protein (HSP) 90 beta in human gastric cancer tissue and SGC7901/VCR of MDR-type gastric cancer cell line. Immunohistochemical staining and in situ hybridization methods. Heat shock protein 90 beta was mainly located in the cell cytoplasma and weakly expressed in non-cancerous gastric mucosa. The expression rates of HSP90 beta in normal gastric mucosa, gastritis and paracancer tissues were 11.76%, 13.04% and 11.42% respectively, and there were no significant differences between them (P > 0.05). The expression of HSP90 beta was increased in gastric cancer. The positive rate of HSP90 beta in gastric cancer tissue was 30.00%, and was higher than non-cancerous gastric mucosa (P < 0.05). The expression rates of HSP90 beta in well differentiated, moderately differentiated, poorly differentiated gastric cancer and mucinous carcinoma were 15.38%, 31.25%, 33.33%, and 42.85% respectively. The expression of HSP90 beta in SGC7901/VCR of MDR-type gastric cell line was higher than in its parental cell line SGC7901. In situ hybridization showed that the positive signal of HSP90 beta was mainly located in the cell cytoplasma. The expression of HSP90 beta was higher in gastric cancer tissue than in non-cancerous gastric mucosa. In gastric cancer tissue, the expression of HSP90 beta was greater in poorly differentiated cancer tissue, and in SGC7901/VCR of MDR-type gastric cancer cell line the expression of HSP90 beta was higher than that in its parental cell line SGC7901.

Significant correlation between expression of heat shock proteins 27, 70 and lymphocyte infiltration in esophageal squamous cell carcinoma

Article

Apr 2002
CANCER LETT

The objective of this study was to clarify the clinicopathologic and prognostic significance of heat shock proteins (HSP) 27 and 70 expression in esophageal squamous cell carcinoma (SCC). Immunohistochemical staining for HSPs 27 and 70 was performed on surgical specimens obtained from 62 patients with esophageal SCC. The expression of both HSPs 27 and 70 correlated inversely with depth of invasion (P<0.05) and pathologic stage (P<0.05), and correlated positively with lymphocyte infiltration (P<0.05). Reduction of HSP 70 expression was significantly correlated with poor prognosis (P<0.05). Patients with HSP 27-negative tumors tended to have a poor prognosis compared with patients with HSP 27-positive tumors. The present findings suggest that HSPs 27 and 70 are significant prognostic factors for esophageal SCC.

Heat-Shock Protein 90: Potential Involvement in the Pathogenesis of Malignancy and Pharmacological Intervention

Article

Nov 2002

Heat-shock protein 90 (Hsp90) is an essential, cytosolic protein. Its overexpression in a wide variety of malignant tumors makes it a candidate target for pharmacological intervention. The association with Hsp90 stabilizes key regulatory proteins like Fak, Bcr-Abl, ErbB2, mutant p53 and Raf-1. The disruption of these heterocomplexes by Hsp90 inhibitors causes the rapid degradation of Hsp90-client proteins by the proteasome. Benzoquinone ansamycins were the first group of compounds for which interference with Hsp90 function was shown to be the major mechanism of action. They are in the early phase of clinical development. Radicicol and its derivatives are functional analogues of benzoquinone ansamycins without structural similarity. Flavonoids and stresgenin B share the ability to suppress heat-shock protein synthesis. Recently, it became apparent that coumarin antibiotics, cisplatin and paclitaxel also bind to Hsp90. The clinical value of the newly characterized agents with activity towards Hsp90 remains to be determined.

Cells in stress: transcriptional activation of heat shock genes

Jan 1993
1409-1410

Ri Marimoto

Marimoto RI. Cells in stress: transcriptional activation of heat shock genes. Science 1993, 259, 1409–1410.

Abundance of heat shock protein (hsp 89, hsp 60 and hsp 27) in malignant cells of HodgkinÕs disease

Jan 1998
5507-5513

Hsu Pl
Hsu
Sm

Hsu PL, Hsu SM. Abundance of heat shock protein (hsp 89, hsp 60 and hsp 27) in malignant cells of HodgkinÕs disease. Cancer Res 1998, 58, 5507–5513.

Esophageal cancer trends and risk factor

Jan 1994
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Blot

Blot WJ. Esophageal cancer trends and risk factor. Semin Oncol 1994, 121, 403–410.

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Jan 2000
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Masao O, Zenya N, Shigeo T, Yuhkinchi M, Goro A. Over expression and localization of heat shock proteins mRNA in pancreatic carcinoma. J Nippon Med Sch 2000, 67, 177–185.

International union against cancer TNM classification of malignant tumors

Jan 1997

Sobin Lh
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Sobin LH, Wittenkid CH, eds. International union against cancer TNM classification of malignant tumors. 5th ed. New York, Wi-ley, 1997.

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Jan 1992
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M Ferrarini
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Ferrarini M, Helatai S, Zocchi MR, Rugarli C. Unusual expres-sion and localization of heat shock proteins in human tumor cells. Int J Cancer 1992, 51, 613–619.

Clinical and biological significance of heat shock protein 89α in human breast cancer

Jan 1992
409

Luqmani

Unusual expression and localization of heat shock proteins in human tumor cells

Jan 1992
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Ferrarini

High constitutive expression of heat shock protein 90α in human acute leukaemia

Jan 1992
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CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation

Jan 2001
42938

Jiang

Negative regulation of cytochrome c-mediated oligomerization of Apaf-1 and activation of procaspase-9 by heat shock protein 90

Jan 2000
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Pandey

Expression of heat-shock protein Hsp60 correlated with the apoptotic index and patient prognosis in human oesophageal squamous cell carcinoma

Abstract

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