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Kai Zhu,
Qi Pan,
Xin Zhang,
Ling-Qun Kong, Jia Fan,
Zhi Dai,
Lu Wang,
Xin-Rong Yang,
Jie Hu,
Jin-Liang Wan,
Yi-Ming Zhao,
Zhong-Hua Tao,
Zong-Tao Chai,
Hai-Ying Zeng,
Zhao-You Tang,
Hui-Chuan Sun,
Jian Zhou
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ABSTRACT: Endothelial cells are critical for angiogenesis, and microRNA play important roles in this process. We investigated the regulatory role of microRNAs in endothelial cells of hepatocellular carcinoma (HCC) by examining the microRNA expression profile of human umbilical vein endothelial cells (HUVECs) in the absence or presence of human HCC cells, and identified miR-146a as the most highly up-regulated microRNA. Furthermore, we revealed that miR-146a promoted the expression of platelet-derived growth factor receptor α (PDGFRA) in HUVECs, and this process was mediated by BRCA1. Overexpression of PDGFRA in the ECs of HCC tissues was associated with microvascular invasion, and predicted a poorer prognosis. These results suggest that MiR-146a plays a key role in regulating the angiogenic activity of ECs in HCC through miR-146a-BRCA1-PDGFRA pathway. MiR-146a may emerge as a potential anti-angiogenic target on ECs for HCC therapy.
Carcinogenesis 05/2013; · 5.70 Impact Factor
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ABSTRACT: AIM: Safety and survival were investigated in patients treated according to expaned surgical indications for colorectal hepatic metastases. METHOD: A retrospective analysis of all consecutive patients who underwent resection of colorectal hepatic metastases at Zhongshan Hospital from 2000 to 2010 was conducted. The patients were divided into two groups based on a change in the surgical indications introduced in 2005. Patients in Group I underwent hepatic surgery between 2000 and 2004, and those in Group II between 2005 and 2010. The clinicopathological data and survival rates of both groups were analyzed. RESULTS: There were 530 patients who underwent hepatic surgery between 2000 and 2010. After the expansion of surgical indications, the rate of surgical resection rose from 25.1% to 35.1% (P < 0.05). There was no significant difference in perioperative mortality (2.2% vs. 0.9%) or morbidity (20.9% vs. 29.8%). Recurrence occurred in 27.5% and 36.7%, in Groups I and II and 5-year overall survival was 43% and 49% (not significant). CONCLUSION: Expanding the indications for surgical resection of hepatic metastases increased the resection rate but had no significant effect on survival. This article is protected by copyright. All rights reserved.
Colorectal Disease 05/2013; · 2.93 Impact Factor
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Rui Liao,
Han Wu,
Yong Yi,
Jia-Xing Wang,
Xiao-Yan Cai,
Hong-Wei He,
Yun-Feng Cheng,
Jian Zhou, Jia Fan,
Jian Sun,
Shuang-Jian Qiu
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ABSTRACT: BACKGROUND: Peritumoral activated hepatic stellate cells (HSCs) are versatile myofibroblast-like cells closely related with hepatocellular carcinoma (HCC) progression. So far, comprehensive comparison of gene expression of human HSCs during hepatocarcinogenesis is scanty. Therefore, we identified the phenotypic and genomic characteristics of peritumoral HSCs to explore the valuable information on the prognosis and therapeutic targets of HBV related HCC. METHODS: A tissue microarray containing 224 HBV related HCC patients was used to evaluate the expression of phenotype markers of HSCs including alpha-SMA, glial fibrillary acidic protein (GFAP), desmin, vinculin and vimentin. HSCs and cancer associated myofibroblasts (CAMFs) were isolated from normal, peritumoral human livers and cancer tissues, respectively. Flow cytometry and gene microarray analysis were performed to evaluate the phenotypic changes and gene expression in HCC, respectively. RESULTS: Peritumoral alpha-SMA positive HSCs showed the prognostic value in time to recurrence (TTR) and overall survival (OS) of HCC patients, especially in early recurrence and AFP-normal HCC patients. Expression of GFAP positive HSCs cell lines LX-2 was significantly decreased after stimulation with tumor conditioned medium. Compared with quiescent HSCs, peritumoral HSCs and intratumoral CAMFs expressed considerable up- and down-regulated genes associated with biological process, cellular component, molecular function and signaling pathways involved in fibrogenesis, inflammation and progress of cancer. CONCLUSIONS: Peritumoral activated HSCs displayed prognostic value in HBV related-HCC, and their genomic characteristics could present rational biomarkers for HCC risk and promising therapeutic targets.
Journal of Experimental & Clinical Cancer Research 04/2013; 32(1):22. · 2.15 Impact Factor
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Han Wu,
Pei Chen,
Rui Liao,
Yi-Wei Li,
Yong Yi,
Jia-Xing Wang,
Xiao-Yan Cai,
Hong-Wei He,
Jian-Jun Jin,
Yun-Feng Cheng, Jia Fan,
Jian Sun,
Shuang-Jian Qiu
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ABSTRACT: BACKGROUND AND AIM: Regulatory T cells (Treg) play a vital role in immunosuppressive crosstalk; however, Tregs from different locations lead to different clinical outcomes. Our aim was, therefore, to compare the prevalences and suppressive phenotypes of Tregs in the peripheral blood, peritumor, and intratumor of patients with hepatocellular carcinoma (HCC). METHODS: The frequencies and phenotypes of CD4(+) CD25(+) CD127(low/-) CD49d(-) Tregs in the periphery, peritumor, and intratumor of 78 HCC patients and 12 healthy controls were evaluated by flow cytometry. Treg-cell suppressive activity was determined using an in vitro CD154 expression assay. Tregs from tumor and paired peritumor were then hybridized using an Agilent whole genome oligo microarray, and selected genes were validated by real-time polymerase chain reaction (PCR). Functional analysis of the microarray data was performed using KEGG and Gene Ontology (GO) analyses. RESULTS: Intratumoral Tregs exhibited higher frequencies and more suppressive phenotypic functions than those in peritumor and periphery, whereas there was no difference between the latter two. Functional analysis showed that complement cascades, p53, and glycosylphatidylinositol-anchor biosynthesis pathways were significantly up-regulated in intratumoral Tregs, the salivary secretion pathway was significantly down-regulated in intratumoral Tregs, and immune cells and tumor-immuno related GO terms were significantly affected. CONCLUSIONS: Tregs in different locations exhibited different functional statuses. A higher prevalence and more suppressive phenotype suggested a critical role for intratumoral Tregs in the formation of multicellular immuno-suppressive networks. HCC immunotherapy may be improved, therefore, by specific locational Tregs elimination or suppression.
Journal of Gastroenterology and Hepatology 03/2013; · 2.87 Impact Factor
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Zhong-Hua Tao,
Jin-Liang Wan,
Ling-Yao Zeng,
Lu Xie,
Hui-Chuan Sun,
Lun-Xiu Qin,
Lu Wang,
Jian Zhou,
Zheng-Gang Ren,
Yi-Xue Li, Jia Fan,
Wei-Zhong Wu
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ABSTRACT: MicroRNAs (miRNAs) play a critical role in tumor metastasis. In this study, we identified a set of 32 miRNAs involved in hepatocellular carcinoma (HCC) metastasis. Among them, miR-612 was shown for the first time to have inhibitory effects on HCC proliferation, migration, invasion, and metastasis. AKT2 was verified to be one of the direct targets of miR-612, through which the epithelial-mesenchymal transition (EMT) and metastasis were inhibited. The level of miR-612 in HCC patients was inversely associated with tumor size, stage, EMT, and metastasis. Of particular importance, miR-612 is involved in both the initial and final steps of the metastatic cascade, by suppressing local invasion and distant colonization. The pleiotropic roles of miR-612 in the HCC metastatic cascade suggest that it could be an effective target for both early and advanced HCC.
Journal of Experimental Medicine 03/2013; · 13.85 Impact Factor
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ABSTRACT: This study aimed to identify serum biomarkers for microvascular invasion (MVI) in hepatocellular carcinoma (HCC). MVI is a histological sign of micrometastasis in the liver and is considered as one of the most powerful prognostic factors in HCC. The serum of HCC patients with different vascular invasion statuses was examined by iTRAQ-based proteomic profiling. The expression levels of 24 proteins were associated with the extent of vascular invasion in the pooled samples of 45 HCC cases. Western blot analyses in 90 HCC cases confirmed the correlation of the expression level of paraoxonase 1 (PON1) with the extent of vascular invasion. ELISA assays demonstrated the diagnostic utility of the PON1 level, with the area under curve values of 0.847 and 0.889 for the MVI and gross vascular invasion, respectively, relative to the patients without vascular invasion, in a cohort of 387 additional HCC cases. Immunohistochemistry revealed that PON1 expression in tumor cells was inversely correlated with the extent of vascular invasion in 200 additional HCC cases. In conclusion, using a proteomic approach, we found that serum PON1 was a novel diagnostic biomarker for MVI. The prognostic values of serum PON1 and its possible therapeutic applications are worth further investigation.
Journal of Proteome Research 03/2013; · 5.11 Impact Factor
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Jie Hu,
Zheng Wang,
Chang-Jun Tan,
Bo-Yi Liao,
Xin Zhang,
Min Xu,
Zhi Dai,
Shuang-Jian Qiu,
Xiao-Wu Huang,
Jian Sun,
Qi-Man Sun,
Yi-Feng He,
Kang Song,
Qi Pan,
Ying Wu, Jia Fan,
Jian Zhou
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ABSTRACT: BACKGROUND: Acute rejection (AR) of an organ transplant is a life-threatening complication. Currently, there are few diagnostic biomarkers suitable for clinical application. We aim to determine the potential of plasma microRNAs as biomarkers for AR. METHODS: Using rat orthotopic liver transplantation model and microarrays, we compared the difference in the spectrum and levels of microRNAs in both plasma and grafts between AR rats and control. AR-related plasma microRNAs were selected and validated using real-time quantification polymerase chain reaction. Plasma from AR rats with or without tacrolimus treatment was used for microRNA dynamic monitoring. To clarify the origin of AR-related plasma microRNAs, drug-induced liver damage rat model were performed and in situ hybridization was used to detect and localize the specific microRNA in allografts. RESULTS: We found that plasma miR-122, miR-192, and miR-146a was significantly up-regulated when AR occur (fold change>2; P<0.05) and the elevation could be repressed by immunosuppression. In liver injury rat model, up-regulated plasma miR-122 (fold change=22.126; P=0.002) and miR-192 (fold change=8.833; P<0.001) rather than miR-146a (fold change=1.181; P=0.594) were observed. Further study demonstrated that miR-146a was up-regulated by sixfold in microvesicles isolated from AR plasma, whereas miR-122 and miR-192 showed no distinct change. In situ hybridization revealed that the portal areas of the AR graft were brimming with lymphocytes, which showed highly intense staining for miR-146a. CONCLUSIONS: Our study provides the global fingerprint of plasma microRNAs in AR rats and suggests that plasma miR-122 and miR-192 reflect liver injury, whereas miR-146a may associate with cellular rejection.
Transplantation 03/2013; · 4.00 Impact Factor
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ABSTRACT: Liver transplantation is one of the curative treatments for hepatocellular carcinoma (HCC). However, cancer recurrence and metastasis after liver transplantation are common in some HCC patients with high-risk factors, even within the Milan criteria. It remains unclear whether adjuvant therapy with sorafenib inhibits HCC recurrence and metastasis after liver transplantation. Therefore, we performed orthotopic liver transplantation in an ACI rat model of HCC. Because liver transplantation involves immune rejection and tolerance and it is unknown whether sorafenib influences the immune response, we also investigated the effects of sorafenib on immune balance. In this study, we established an allogeneic rat liver transplantation model with a liver graft from Lewis to ACI rats bearing orthotopic HCC and administrated cyclosporine to prevent acute allograft rejection. From day 7 after liver transplantation, sorafenib was administrated at 30 mg/kg/day for 3 weeks. Our results showed that the serum levels of vascular endothelial growth factor and hepatocyte growth factor significantly increased after liver transplantation, and the Th1/Th2 immune balance was shifted toward a Th2 response after immunosuppressant administration. Compared with controls, sorafenib significantly inhibited ERK phosphorylation, improved progression-free survival and overall survival. The tumor proliferation rate and angiogenesis in the post-transplant recurrent tumor tissues decreased, and the tumor apoptosis rate increased in the sorafenib group. There was no significant difference in the Th1/Th2 immune balance between the sorafenib and control groups. In conclusion, adjuvant therapy with sorafenib is highly effective at inhibiting cancer recurrence and metastasis without influencing the immune balance after liver transplantation for HCC with high expression of pERK. This study suggests that sorafenib may have potential particularly as part of a stratified medicine approach to HCC treatment after liver transplantation. © 2013 American Association for the Study of Liver Diseases.
Liver Transplantation 02/2013; · 3.39 Impact Factor
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Chao Zhao,
Peng Yin,
Chuanzhong Mei,
Na Li,
Wantong Yao,
Xin Li,
Jingjing Qi,
Kun Fan,
Zengxia Li,
Liying Wang,
Yinghong Shi,
Shuangjian Qiu, Jia Fan,
Xiliang Zha
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ABSTRACT: Abnormal DNA methylation is one of the important characteristics in tumor cells. Apoptosis plays an essential role in cell survival and processing. It is not clear whether DNA methyltransferases (DNMTs) change in apoptosis and how DNMTs are regulated in apoptosis. In this study, we found that SMMC-7721 or BEL-7404 cells were induced to apoptosis by STS, meanwhile the DNMT3B protein and mRNA level were decreased. To explore the mechanism of DNMT3B down-regulation, we found that the mRNA decay was not changed and core promoter activity of DNMT3B gene was decreased in STS-induced apoptosis. In order to figure out the signal molecule involved in transcriptional regulation of DNMT3B gene by STS, p-JNK, p-ERK, and p-p38 were examined. In STS-induced apoptosis p-JNK level was increased, and p-ERK and p-p38 were decreased. Furthermore, the inhibitor of p-JNK significantly alleviated the decline of DNMT3B protein. We also found that the siRNA of DNMT3B strengthened the cleavage of PARP and pro-caspase-3 as well as up-regulated the p16 gene expression in STS-treated cells. We concluded here that STS-regulated DNMT3B gene expression via p-JNK and down-regulation of DNMT3B-mediated STS-induced apoptosis through the up-regulation p16 expression.
Molecular and Cellular Biochemistry 02/2013; · 2.06 Impact Factor
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Anuradha Budhu,
Stephanie Roessler,
Xuelian Zhao,
Zhipeng Yu,
Marshonna Forgues,
Junfang Ji,
Edward Karoly,
Lun-Xiu Qin,
Qing-Hai Ye,
Hu-Liang Jia, Jia Fan,
Hui-Chuan Sun,
Zhao-You Tang,
Xin Wei Wang
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ABSTRACT: BACKGROUND & AIMS: We combined gene expression and metabolic profiling analyses to identify factors associated with outcomes of patients with hepatocellular carcinoma (HCC). METHODS: We compared metabolic and gene expression patterns between paired tumor and non-tumor tissues from 30 patients with HCC, and validated the results using samples from 356 patients with HCC. A total of 469 metabolites were measured using liquid chromatography/mass spectrometry and gas chromatography/mass spectrometry. Metabolic and genomic data were integrated, and Kaplan-Meier and Cox proportional hazards analyses were used to associate specific patterns with patient outcomes. Associated factors were evaluated for their effects on cancer cells in vitro and tumor formation in nude mice. RESULTS: We identified 28 metabolites and 169 genes associated with aggressive HCC. Lipid metabolites of stearoyl-CoA-desaturase (SCD) activity were associated with aberrant palmitate signaling in aggressive HCC samples. Expression of gene products associated with these metabolites, including SCD, were independently associated with survival times and tumor recurrence in the test and validation sets. Combined expression of SCD and α-fetoprotein were associated with outcomes of patients with early-stage HCC. Levels of MUPA (monounsaturated palmitic acid), the product of SCD activity, were increased in aggressive HCCs; MUPA increased migration and invasion of cultured HCC cells and colony formation by HCC cells. HCC cells that expressed small interfering RNA against SCD had decreased cell migration and colony formation in culture and reduced tumorigenicity in mice. CONCLUSIONS: Using a combination of gene expression and metabolotic profile analysis, we identified a lipogenic network that involves SCD and palmitate signaling and was associated with HCC progression and patient outcomes.
Gastroenterology 01/2013; · 11.68 Impact Factor
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Xiao-Yong Huang,
Ai-Wu Ke,
Guo-Ming Shi,
Xin Zhang,
Chi Zhang,
Ying-Hong Shi,
Xiao-Ying Wang,
Zhen-Bin Ding,
Yong-Sheng Xiao,
Jun Yan,
Shuang-Jian Qiu, Jia Fan,
Jian Zhou
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ABSTRACT: The overall survival of patients with hepatocellular carcinoma (HCC) remains poor, and the molecular pathogenesis remains incompletely defined in HCC. Here, we report that increased expression of αB-Crystallin in human HCC predicts poor survival and disease recurrence after surgery. Multivariate analysis identifies αB-Crystallin expression as an independent predictor for postoperative recurrence and overall survival. We unveil that elevated expression of αB-Crystallin promotes HCC progression in vivo and in vitro. We demonstrate that αB-Crystallin overexpression fosters HCC progression by inducing epithelial-mesenchymal transition (EMT) in HCC cells through activation of the ERK cascade, which can counteract the effect of sorafenib. αB-Crystallin complexes with and elevates 14-3-3ζ protein, leading to up-regulation of ERK1/2 activity. Moreover, overexpression of αB-Crystallin in HCC cells induces EMT progression through an ERK1/2/Fra-1/slug signaling pathway. Clinically, our data reveal that overexpression of both αB-Crystallin and 14-3-3ζ correlates with the HCC poorest survival outcomes, and sorafenib response is impaired in patients with αB-Crystallin overexpression. Conclusions: These data suggest that the αB-Crystallin-14-3-3ζ complex acts synergistically to promote HCC progression by constitutively activating ERK signaling. This study reveals αB-Crystallin as a potential therapeutic target for HCC and a biomarker for predicting sorafenib treatment response. (HEPATOLOGY 2013.).
Hepatology 01/2013; · 11.66 Impact Factor
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Rui Liao,
Jian Sun,
Han Wu,
Yong Yi,
Jia-Xing Wang,
Hong-Wei He,
Xiao-Yan Cai,
Jian Zhou,
Yun-Feng Cheng, Jia Fan,
Shuang-Jian Qiu
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ABSTRACT: BACKGROUND: Hepatocellular carcinoma (HCC) is a typical malignancy in a background of chronic inflammation. Th17 cells (a major source of IL-17) constitute crucial components of infiltrating inflammatory/immune cells in HCC and can amplify inflammatory response via binding to interleukin-17 receptor (IL-17R). Thus, we investigated the expression and clinical significance of IL-17 and IL-17 receptor family cytokines in HCC. METHODS: The expression and prognostic value of IL-17 and IL-17R (A-E) were examined in 300 HCC patients after resection. Six Th17 associated cytokines in serum (n = 111) were quantified using enzyme-linked immunosorbent assays. Phenotypic features of IL-17+ CD4+ T cells were determined by flow cytometry analysis. RESULTS: High expression of intratumoral IL-17 and IL-17RE were significantly associated with poorer survival (p = 0.016 and <0.001, respectively) and increased recurrence (both P < 0.001) of HCC patients. Moreover, intratumoral IL-17, individually or synergistically with IL-17RE, could predict HCC early recurrence and late recurrence. Also, peritumoral IL-17RE showed the prognostic ability in HCC (P < 0.001 for OS/TTR). Furthermore, expression levels of Th17 associated cytokines including IL-6, -22, -17R and TNF-alpha were increased in serum of HCC patients compared to haemangioma patients. Importantly, activated human hepatic stellate cells induced in vitro expansion of IL-17+ CD4+ T cells. CONCLUSIONS: High expression of IL-17 and IL-17RE were promising predictors for poor outcome of HCC patients. The protumor power of IL-17 producing CD4+ T cells was probably involved in the crosstalk with different types of inflammatory/immune cells in HCC.
Journal of Experimental & Clinical Cancer Research 01/2013; 32(1):3. · 2.15 Impact Factor
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ABSTRACT: PURPOSE: The biology underlying bone-specific metastasis (BM) of hepatocellular carcinoma (HCC) is poorly understood. The goal of the present study is to further elucidate the molecular and cellular mechanisms underlying HCC with BM. METHODS: The expression of connective tissue growth factor (CTGF) and interleukin-11 (IL-11) in RNA extracted from 127 formalin-fixed, paraffin-embedded HCC specimens was examined by quantitative real-time polymerase chain reaction. A cellular hypoxic model was established in vitro to investigate CTGF and osteoprotegerin (OPG) expression and roles in hypoxia-induced tumor aggressiveness. RESULTS: The mean CTGF expression in BM versus non-metastatic samples was 3.63-times higher, and IL-11 expression was detected in 62.5 % (10/16) of BM samples versus only in 18.9 % (21/111) of the non-metastatic ones. Highly metastatic HCC cell lines tended to show strong expression of CTGF and IL-11, but low expression of OPG. Hypoxic stimulation of HCC 97L cells increased the level of CTGF mRNA by 2.80-fold within 1.5 h, and hypoxia-inducible factor-1α mRNA levels in these cells could be increased by stimulation with recombinant CTGF protein. Furthermore, OPG and matrix metalloproteinase-2 and -9 levels were also induced under hypoxic conditions. CONCLUSIONS: Expression levels of intratumoral CTGF or IL-11 were independent prognostic factors for the development of BM in HCC patients. Tumor hypoxia enhanced the expression of CTGF, which initiates the invasive angiogenesis cascade and enhances expression of many hypoxia-associated genes. Cellular release of OPG may play a role in tumor cell survival. The hypoxia-induced cascade in HCC cells may contribute to invasion and metastasis in vivo.
Journal of Cancer Research and Clinical Oncology 01/2013; · 2.56 Impact Factor
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ABSTRACT: Calpain small subunit 1 (Capn4) has been shown to correlate with the metastasis/invasion of hepatocellular carcinoma. This study aimed to investigate the role of Capn4 in intrahepatic cholangiocarcinoma (ICC).
Capn4 expression was measured in 33 ICC tissues by quantitative real-time polymerase chain reaction and western blot. The role of Capn4 in the migration, invasion and proliferation of ICC cells and matrix metalloproteinase 2 (MMP2) expression were assessed after Capn4 depletion by specific small interfering RNA. Capn4 expression was further examined by immunohistochemistry in a tissue microarray consisting of 140 ICC patients and 13 normal liver tissues, and the prognostic role of Capn4 in ICC was evaluated by Kaplan-Meier and Cox regression analyses.
Capn4 expression was significantly higher in the ICC tissues compared to the peritumor tissues. Capn4 down-regulation impaired the migration/invasion ability of HCCC-9810 and QBC939 cells in vitro and decreased MMP2 expression. Capn4 overexpression significantly correlated with the presence of lymphatic metastasis of ICC (p = 0.026) and the tumor-node-metastasis (TNM) stage (p = 0.009). The postoperative 2- and 5-year overall survivals in patients with Capn4(low) were higher than those in the Capn4(high) group. The cumulative recurrence rate in patients with Capn4(low) was much lower than in the Capn4(high) group. Multivariate analysis showed that Capn4 overexpression was an independent prognostic marker in ICC.
Capn4 overexpression was implicated in ICC metastasis/invasion, and Capn4 overexpression may be used as a molecular therapeutic target for ICC.
PLoS ONE 01/2013; 8(1):e54619. · 4.09 Impact Factor
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ABSTRACT: BACKGROUD: RNA interference (RNAi) has recently emerged as a potential treatment modality for hepatocellular carcinoma (HCC) therapy, but the lack of cellular targets and sustained efficacy limits its application. The purpose of this study is to develop an HCC tissue-specific RNAi system and investigate its possibility for HCC treatment.
Two different HCC-specific RNAi systems in which therapeutic miRNA or shRNA against target gene (Beclin 1) was directly or indirectly driven by alpha-fetoprotein promoter (AFP-miRNA and AFP-Cre/LoxP-shRNA) were constructed. Human HCC cell lines (HepG2, Hep3B and HCCLM3) and non-HCC cell lines (L-02, Hela and SW1116) were infected with the systems. The effectiveness and tissue-specificity of the systems were examined by Q-PCR and western blot analysis. The efficacy of the systems was further tested in mouse model of HCC by intravenous or intratumoral administration. The feasibility of the system for HCC treatment was evaluated by applying the system as adjuvant therapy to enhance sorafenib treatment. An AFP-Cre/LoxP-shRNA system targeting Atg5 gene (AFP-Cre/LoxP-shRNA-Atg5) was constructed and its efficacy in sensitizing HCC cells (MHCC97L/PLC) to sorafenib treatment was examined by apoptosis assay and tumorigenesis assay .
The AFP-miRNA system could silence target gene (Beclin 1) but required a high titer which was lethal to target cells. The AFP-Cre/LoxP-shRNA system could efficiently knockdown target gene while maintain high HCC specificity. Intratumoral injection of the AFP-Cre/LoxP-shRNA system could efficiently silence target gene (Beclin 1) while intravenous administration could not. The AFP-Cre/LoxP-shRNA system target Atg5 gene could significantly sensitize MHCC97L/PLC cells to sorafenib-induced apoptosis and tumor growth suppression .
An efficient HCC tissue-specific RNAi system (AFP-Cre/LoxP-shRNA) was successfully established. The system provides a usable tool for HCC-specific RNAi therapy, which may serve as a new treatment modality for HCC.
PLoS ONE 01/2013; 8(2):e53072. · 4.09 Impact Factor
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Ling-Qun Kong,
Xiao-Dong Zhu,
Hua-Xiang Xu,
Ju-Bo Zhang,
Lu Lu,
Wen-Quan Wang,
Qiang-Bo Zhang,
Wei-Zhong Wu,
Lu Wang, Jia Fan,
Zhao-You Tang,
Hui-Chuan Sun
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ABSTRACT: Our previous study has found that the abundance of peritumoral CD68(+) macrophages was associated with poor prognosis in hepatocellular carcinoma (HCC) after resection. However, CD68 staining could not discriminate the protumoral or tumoricidal subpopulations from pan-macrophages. CD163 is a marker of alternatively activated macrophages. In this study, the clinical significance of CD163(+) cells in tumors and peritumoral liver tissues was evaluated in a cohort of 295 patients with HCC after curative resection. We found that the density of CD163(+) cells was well correlated with that of CD68(+) cells in both tumors and peritumoral liver tissues but was much more. Immunostaining on consecutive sections and flow cytometry assay on surgical resected specimens further supported the findings that the CD163(+) cells was more abundant than CD68(+) cells. The density of peritumoral CD68(+) cells was associated with poor recurrence-free survival (RFS) and poor overall survival (OS) (P = 0.004 and P = 0.001, respectively), whereas the CD163(+) cells have no prognostic values either in tumors or in peritumoral liver tissues. In another cohort of 107 HCC patients, preoperative plasma concentration of soluble form of CD163 (sCD163) was associated with active hepatitis-related factors but not associated with the markers of tumor invasion. In conclusion, both the CD163(+) cells local infiltration and plasma sCD163 were of limited significance in HCC, and they were more likely markers related to active hepatitis rather than tumor progression.
PLoS ONE 01/2013; 8(3):e59771. · 4.09 Impact Factor
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Junfang Ji,
Lei Yu,
Zhipeng Yu,
Marshonna Forgues,
Takahiro Uenishi,
Shoji Kubo,
Kenichi Wakasa,
Jian Zhou, Jia Fan,
Zhao-You Tang,
Shijun Fu,
Hongguang Zhu,
Jason Gang Jin,
Hui-Chuan Sun,
Xin Wei Wang
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ABSTRACT: Background & Aims: Adjuvant therapies for hepatocellular carcinoma (HCC) such as interferon-alpha are effective only in a subset of patients. Previously we found that HCC patients with low level of miR-26 have survival benefits from interferon-alpha. The purpose of this study is to develop a standardized miR-26 diagnostic test (referred as MIR26-DX) to assist identification of candidate HCC patients for adjuvant interferon-alpha therapy. Methods: We developed a multiplex reverse-transcription quantitative polymerase-chain-reaction assay to determine the levels of two HCC-related miR-26 transcripts along with six small RNA reference transcripts. We evaluated archived paraffin-embedded tissues from three cohorts of HCC patients (n=248) who underwent radical resection at three different clinical centers. Fifty-two percent of them underwent adjuvant interferon-alpha therapy. We used Cox-Mantel log-rank test to evaluate patient survival. Results: We found that the multiplexing assay was stable and reproducible regardless of differences in sample preparations and operators. We developed a matrix template and a scoring algorithm based on a training cohort (n=129) to assign HCC patients, and then applied the template in two test cohorts (n=119). The proportions of HCC patients assigned as low miR-26 by this algorithm were 68, 4, and 63 percent in the training cohort and two test cohorts, respectively. Consistently, HCC with low miR-26 had a favorable response to interferon-alpha with improved median overall survival (≥3year). Conclusions: MIR26-DX is a simple and reliable companion diagnostic test to select HCC patients for adjuvant interferon-alpha therapy.
International journal of biological sciences 01/2013; 9(3):303-12. · 2.70 Impact Factor
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Zhi-Chao Wang,
Qiang Gao,
Jie-Yi Shi,
Liu-Xiao Yang,
Jian Zhou,
Xiao-Ying Wang,
Ying-Hong Shi,
Ai-Wu Ke,
Guo-Ming Shi,
Zhen-Bin Ding,
Zhi Dai,
Shuang-Jian Qiu, Jia Fan
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ABSTRACT: BACKGROUND: Frequent deletions of the kinesin-like protein gene 1B (KIF1B) have been reported in neural tumors. Recently, a genome-wide association study revealed an association between polymorphisms in the KIF1B gene and the risk of hepatocellular carcinoma (HCC), and several case-control studies have further investigated this relationship. However, these studies have yielded controversial results. We therefore performed a meta-analysis to derive a more precise estimation of the association between the KIF1B gene polymorphisms and HCC risk. METHODOLOGYPRINCIPAL FINDING: PubMed, EMBASE, the ISI Web of Science and the CNKI databases were systematically searched to identify relevant studies. A total of 5 studies containing 13 cohorts with 5,773 cases and 6,404 controls were included. Odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were used to assess the strength of the associations. Subgroup analyses were conducted based on ethnicities, sample sizes and quality scores. Overall, the G allele at rs17401966 of the KIF1B gene was associated with a significantly decreased risk for HCC (OR = 0.81, 95%CI: 0.70-0.93; P = 0.003). Furthermore, subgroup analyses showed that the G allele at rs17401966 of the KIF1B gene significantly reduced the risk for HCC in Chinese cohorts (OR = 0.76, 95%CI: 0.64-0.90; P = 0.002), large-sample-size cohorts (OR = 0.80, 95%CI: 0.73-0.88, P<0.01) and high-quality cohorts (OR = 0.78, 95%CI: 0.71-0.87, P<0.01). However, no significant associations were found in small-sample-size cohorts, studies with low-quality scores and when excluding the cohorts from the study reporting the original discovery. CONCLUSIONSIGNIFICANCE: These findings demonstrate that the presence of the G allele at rs17401966 of the KIF1B gene may decrease the risk for HCC and suggest that KIF1B may play a critical role in the development of HCC. High-quality studies with larger sample sizes and different ethnic populations will be of great value to further confirm these findings.
PLoS ONE 01/2013; 8(4):e62571. · 4.09 Impact Factor
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ABSTRACT: BACKGROUND & AIMS: The immunosuppressive network within the tumor microenvironment is one of the major obstacles to the success of cancer immunotherapy. γδ T cells are attractive effectors for cancer immunotherapy. Nevertheless, the promising anti-tumor effect in vitro is partially if not totally mitigated in vivo. Thus, understanding the immune status of tumor-infiltrating γδ T cells is essential for orchestrating effective immunotherapy strategies. In this study we have investigated the immunophenotype and function of γδ T cells in hepatocellular carcinoma (HCC) patients. METHODS: The phenotype of γδ T cells in peripheral blood, and peritumoral and tumoral tissues of HCC patients (n = 61) was characterized by flow cytometry. Functional analysis of the HCC-infiltrating γδ T cells was conducted directly after γδ T cell isolation. RESULTS: The infiltration of γδT cells in tumoral tissues was significantly reduced compared to paired peritumoral tissues. Impairment in degranulation of the granule pathway and down-regulation of IFN-γ secretion were also demonstrated in HCC-infiltrating γδT cells, which was in agreement with the results of the gene microarray analysis, and further strengthened by the compromised specific cytotoxicity and IFN-γ secretion in vitro. Moreover, isolated HCC-infiltrating CD4(+)CD25(+) regulatory T cells (Treg cells) directly suppressed the cytotoxic function and IFN-γ secretion of γδT cells in a TGFβ- and IL-10-dependent manner. CONCLUSIONS: The effector function of γδT cells was substantially impaired in HCC, which is partially mediated by Treg cells. We propose a new mechanism by which immune privilege develops within the tumor milieu.
Journal of Hepatology 12/2012; · 9.26 Impact Factor
