[Show abstract][Hide abstract] ABSTRACT: Introduction
It is postulated that focal IRE affords complete ablation of soft-tissue tumours while protecting the healthy peritumoral tissue. Therefore, IRE may be an interesting option for minimally invasive, kidney-tissue-sparing, non-thermal ablation of renal tumours.
With this current pilot study (“IRENE trial”), we present the first detailed histopathological data of IRE of human RCC followed by delayed tumour resection. The aim of this interim analysis of the first three patients was to investigate the ablation efficiency of percutaneous image-guided focal IRE in RCC, to assess whether a complete ablation of T1a RCC and tissue preservation with the NanoKnife system is possible and to decide whether the ablation parameters need to be altered.
Following resection 4 weeks after percutaneous IRE, the success of ablation and detailed histopathological description were used to check the ablation parameters.
The IRE led to a high degree of damage to the renal tumours (1 central, 2 peripheral; size range 15–17 mm). The postulated homogeneous, isomorphic damage was only partly confirmed. We found a zonal structuring of the ablation zone, negative margins and, enclosed within the ablation zone, very small tumour residues of unclear malignancy.
According to these initial, preliminary study results of the first three renal cases, a new zonal distribution of IRE damage was described and the curative intended, renal saving focal ablation of localised RCC below <3 cm by percutaneous IRE by the NanoKnife system appears to be possible, but needs further, systematic evaluation for this treatment method and treatment protocol.
CardioVascular and Interventional Radiology 09/2015; DOI:10.1007/s00270-015-1200-6 · 2.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background:
The dicarbonyls glyoxal and methylglyoxal are reactive alde-
hydes with significant toxicity towards many cell types. Glyoxalase 1 is a
major defence enzyme against dicarbonyl stress and upregulated in many
cancer entities. We have recently shown that tamoxifen resistant mamma
carcinoma cells derived from the estrogen receptor positive MCF-7 cell
line exhibited increased sensitivity towards these aldehydes, which corre-
lated with reduced amounts of free sulfhydryl groups in the tamoxifen re-
sistant cell line. Here, we analysed whether glyoxalase 1 is responsible for
aldehyde defence in these cells.
Glyoxalase (GLO1) expression was determined by qRT-PCR
and Western-Blotting. Reactive oxygen species (ROS) were determined by
the ROS sensitive dye 2',7'-dichlorodihydrofluorescein diacetate (H2DCF-
DA) and cell viability/proliferation was assessed by the resazurin assay. A
specific glyoxalase 1 inhibitor and siRNA technique was applied to mod-
ulate GLO1 activity. The inhibitors SB203580 and UO126 were used to
study the importance of MAP-kinase activity and diphenylene-iodonium
(DPI) was applied as specific inhibitor of NADPH-oxidases.
Aldehyde stress induced the production of reactive oxygen spe-
cies by NADPH-oxidase in MCF-7 cells as well as in a tamoxifen resistant
derivative (TamR). Inhibition of p38 MAPK caused reduced production
of ROS. Inhibition of GLO1 resulted in higher sensitivity towards dicar-
bonyl stress. GLO1-protein expression was not altered in the tamoxifen
resistant cells, but the expression of the DJ-1 glyoxalase was significant-
ly reduced. Also, mRNA-accumulation of several NF-kB subunits was al-
tered in the TamR cell line, which correlates with the observation that
TamR but not MCF-7 cells responded to aldehyde stress by phosphoryla-
tion of IkBalpha and this further supports the proposal that NF-kB activ-
ity contributes to tamoxifen resistance.
Expression of glyoxalase 1 and DJ-1 might be useful as prog-
nostic markers in ER-positive mamma carcinomas.
99. Jahrestagung der Deutschen Gesellschaft für Pathologie e.V., Frankfurt a. M. , Germany; 05/2015
[Show abstract][Hide abstract] ABSTRACT: Adenocarcinomas at the gastro-oesophageal junction (GOJ) are currently stratified by tumour location. This retrospective study examines the association of preneoplastic conditions and inflammation of the gastric mucosa with GOJ cancer at different locations and compares them with nonjunctional gastric cancers.
A total of 520 patients with junctional and nonjunctional gastric cancer were assessed for the presence and degree of intestinal metaplasia, glandular atrophy and inflammation in the stomach. Histopathological data were complete for 428 patients (68.9% men, median age 67.7 years), including 172 patients with GOJ cancer (GOJ1: 1-5 cm proximal to the junction, GOJ2: 'true' junctional, GOJ3: 2-5 cm distal to the junction). Gastric inflammation and preneoplastic conditions were scored according to the updated Sydney classification and further stratified into respective operative link on gastritis assessment (OLGA) and operative link on gastritis assessment on intestinal metaplasia (OLGIM) stages.
The prevalence and degree of gastric atrophy and intestinal metaplasia were significantly lower in GOJ1 than GOJ3 (P<0.01). Preneoplastic conditions in the stomach were similar in GOJ3 compared with nonjunctional gastric cancer. GOJ1 were almost exclusively (98.4%) of the intestinal type, whereas GOJ2 and GOJ3 were the diffuse type in 22.6 and 22.4% of the patients (P<0.001). Of all patients, only 8.5 and 12.7% presented with stage III/IV according to OLGA and OLGIM, respectively. However, data for OLGA and OLGIM staging were only available in 61.2 and 67.9% of patients, respectively.
GOJ1 are less likely to be associated with gastric pathology compared with GOJ3 or nonjunctional gastric cancer. OLGA or OLGIM staging in patients with advanced gastro-oesophageal cancer seems to be of limited value.
European journal of gastroenterology & hepatology 05/2015; 27(5):492-500. DOI:10.1097/MEG.0000000000000299 · 2.25 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The lysosomal cysteine carboxypeptidase cathepsin X (CTSX), localized predominantly in immune cells, has been associated with the development and progression of cancer. To determine its specific role in colorectal carcinoma (CRC), we analyzed CTSX expression in non-malignant mucosa and carcinoma of 177 patients as well as in 111 adenomas and related it with clinicopathological parameters. Further, the role of CTSX in the adhesion and invasion of the colon carcinoma cell lines HT-29 and HCT116 was investigated in an in vitro culture cell system with fibroblasts and monocytes, reflecting the situation at the tumor invasion front.
Epithelial CTSX expression significantly increased from normal mucosa to adenoma and carcinoma, with highest expression levels in high grade intraepithelial neoplasia and in early tumor stages. Loss of CTSX occurred with tumor progression, and correlated with advanced local invasion, lymph node and distal metastasis, lymphatic vessel and vein invasion, tumor cell budding and poorer overall survival of patients with CRC. The subcellular distribution of CTSX changed from vesicular paranuclear expression in the tumor center to submembranous expression in cells of the invasion front. Peritumoral macrophages showed highest expression of CTSX. In vitro assays identified CTSX as relevant factor for cell-cell adhesion and tumor cell anchorage to fibroblasts and basal membrane components, whereas inhibition of CTSX caused increased invasiveness of colon carcinoma cells in mono- and co-culture. In conclusion, CTSX is involved in early tumorigenesis and in the stabilization of tumor cell formation in CRC. The results suggest that loss of CTSX may be needed for tumor cell detachment, local invasion and tumor progression. In addition, CTSX in tumor-associated macrophages indicates a role for CTSX in the anti-tumor immune response.
Pathology - Research and Practice 12/2014; 210(12). DOI:10.1016/j.prp.2014.08.014 · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Cathepsin X (CTSX, also called cathepsin Z/P) is a cysteine protease that still plays an unknown role in human cancer. It has been shown to bind cell surface heparin sulphate proteoglycans and integrins, indicating possible functions of CTSX in cellular adhesion, phagocytosis, and immune response. Our previous studies have shown an association between Helicobacter pylori (H. pylori) infection, a strong up-regulation of CTSX, and development of gastric cancer. In this study, yeast two-hybrid analysis revealed that RPLP0, a ribosomal protein P0, interacts with the human CTSX protein in gastric cancer. The CTSX/RPLP0 interaction was confirmed by co-immunoprecipitation assays. In addition, co-localization studies in cancer cell line N87 and gastric cancer tissue samples were performed. Laserscan microscopy revealed a shuttling of RPLP0 (and CTSX) from cytoplasm to the nucleus after CTSX knockdown. Down-regulation of RPLP0 resulted in G1 arrest of gastric cancer cells, whereas knockdown of CTSX led to G1 arrest and apoptosis after 48 h. Knockdown of both proteins caused increased apoptosis. RPLP0 deficiency could suppress cell growth and cell cycle progression by down-regulating CDK2. It was further demonstrated that RPLP0 affected p21 expression, but did not change the expression of Cyclin E. Down-regulation of both proteins at least through CDK2 suggests an anti-apoptotic effect on gastric cancer cells and opens up new possibilities for apoptotic immune modulation and gastric cancer therapy.
Pathology - Research and Practice 09/2014; 211(1). DOI:10.1016/j.prp.2014.09.005 · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective
Interleukin (IL)-1 signalling plays an important role in inflammatory and also malignant processes. IL-1 signals essentially via nuclear factor (NF)-κB, which controls the expression of genes involved in cell proliferation, invasion, angiogenesis and metastasis, among them vascular endothelial growth factor A (VEGF-A). As microenvironment-derived IL-1β is required for invasion and angiogenesis in malignant tumours, such as chondrosarcomas, we investigated whether the anti-inflammatory compound curcumin is able to interfere with IL-1-induced VEGF-a secretion.
Cell cultures of chondrosarcoma cell lines C3842 and SW1353 were treated with IL-1ß and curcumin and VEGF-A secretion was analysed by Western blotting. Gene expression was determined by qRT-PCR. The angiogenenic potential of cell culture supernatants was investigated by a 2-dimensional angiogenesis assay using human umbilical chord endothelial cells (HUVECs). Activation of the nuclear factor NF-κB was analysed by determination of IκBα phosphorylation on Western blots and immunofluorescence.
IL-1 signalling and VEGF-A expression are blocked by curcumin in chondrosarcoma cells. We further show that curcumin blocks IL-1β-induced angiogenesis and NF-κB-related gene expression.
We propose that IL-1 blockade is an additionally treatment option in chondrosarcoma, either by curcumin, its derivatives or other IL-1 blocking agents.
26th European Congress of Pathology, London, UK; 09/2014
[Show abstract][Hide abstract] ABSTRACT: Tamoxifen is the standard adjuvant endocrine therapy for estrogen-receptor positive premenopausal breast cancer patients. However, tamoxifen resistance is frequently observed under therapy. A tamoxifen resistant cell line has been generated from the estrogen receptor positive mamma carcinoma cell line MCF-7 and was analyzed for putative differences in the aldehyde defence system and accumulation of advanced glycation end products (AGE). In comparison to wt MCF-7 cells, these tamoxifen resistant cells were more sensitive to the dicarbonyl compounds glyoxal and methylglyoxal and displayed increased caspase activity, p38-MAPK- and IκBα-phosphorylation. However, mRNA accumulation of the aldehyde- and AGE-defence enzymes glyoxalase-1 and -2 (GLO1, GLO2) as well as fructosamine-3-kinase (FN3K) was not significantly altered. Tamoxifen resistant cells contained less free sulfhydryl-groups (glutathione) suggesting that the increased sensitivity towards the dicarbonyls was due to a higher sensitivity towards reactive oxygen species which are associated with dicarbonyl stress. To further analyse, if these data are of more general importance, key experiments were replicated with tamoxifen resistant MCF-7 cell lines from two independent sources. These cell lines were also more sensitive to aldehydes, especially glyoxal, but were different in their cellular signalling responses to the aldehydes. In conclusion, glyoxalases and other aldehyde defence enzymes might represent a promising target for the therapy of tamoxifen resistant breast cancers.
PLoS ONE 07/2014; 9(7):e101473. DOI:10.1371/journal.pone.0101473 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aims. The alpha-oxo-aldehydes glyoxal and methylglyoxal are byproducts
of fatty acid oxidation and glycolysis. Due to the high glycolytic
activity caused by the Warburg effect, cancer cells produce significantly
more aldehydes than other cells. These substances are highly reactive
and modify amino groups on proteins, often resulting in loss of function.
These modifications also result in stable accumulating products,
the so called advanced glycation end products (AGEs). In this study we
analysed the responses of the estrogen receptor positive mamma carcinoma
cell line MCF-7 to glyoxal and methylglyoxal to evaluate if dialdehyde
defence might be a target for breast cancer treatment.
Methods. Cell vitality was determined by the resazurin assay. Activation
of MAP-kinases and NF-κB was determined by Western blotting,
using specific antibodies against the phosphorylated proteins. Oxidative
stress was measured by loading the cells with the oxidation-sensitive
fluorochrome dihydrofluorescein-diacetate. Mode of cell death was
analysed by caspase activity assay and annexin-V flow cytometry.
Results. Dialdehydes resulted in cell death in low millimolar concentrations.
Cells exposed to the aldehydes responded by a rapid activation
of MAP-kinase signalling especially p38-MAPK and production of reactive
oxygen species as well as activation of caspase 3/7 activity, finally
leading to cell death. Interestingly, a tamoxifen resistant derivative of
MCF-7 showed an additional activation of the NF-κB transcription factor
by methylglyoxal only and was altogether more sensitive to the aldehydes. Addition of the antioxidant N-acetyl cystein protected the cells,
demonstrating that oxidative stress was a major cause of cell death.
Conclusions. These data demonstrate that exogenous aldehyde stress is
cytotoxic to mamma carcinoma cells by causing severe oxidative stress.
Inhibition of aldehyde defence enzymes such as glyoxalases might
therefore be a promising therapeutic option against breast cancer.
98. Jahrestagung der Deutschen Gesellschaft für Pathologie e. V., Berlin, Germany; 06/2014
[Show abstract][Hide abstract] ABSTRACT: Interleukin (IL)-1 signaling plays an important role in inflammatory processes, but also in malignant processes. The essential downstream event in IL-1 signaling is the activation of nuclear factor (NF)-κB, which leads to the expression of several genes that are involved in cell proliferation, invasion, angiogenesis and metastasis, among them VEGF-A. As microenvironment-derived IL-1β is required for invasion and angiogenesis in malignant tumors, also in chondrosarcomas, we investigated IL-1β-induced signal transduction and VEGF-A expression in C3842 and SW1353 chondrosarcoma cells. We additionally performed in vitro angiogenesis assays and NF-κB-related gene expression analyses. Curcumin is a substance which inhibits IL-1 signaling very early by preventing the recruitment of IL-1 receptor associated kinase (IRAK) to the IL-1 receptor. We demonstrate that IL-1 signaling and VEGF-A expression are blocked by Curcumin in chondrosarcoma cells. We further show that Curcumin blocks IL-1β-induced angiogenesis and NF-κB-related gene expression. We suppose that IL-1 blockade is an additional treatment option in chondrosarcoma, either by Curcumin, its derivatives or other IL-1 blocking agents.
PLoS ONE 06/2014; 9(6):e99296. DOI:10.1371/journal.pone.0099296 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of the current study was to investigate the role of BRCA1 promoter methylation as predictive factor of response to platinum-taxane-based therapy in sporadic ovarian cancer.
BRCA1 promoter methylation was analyzed in 42 sporadic epithelial ovarian cancers. The results were validated in a second cohort of 137 ovarian cancer patients.
BRCA1 promoter methylation was observed in 35.7 % of patients in the first group and in 33.6 % in the second group. BRCA1 promoter methylation was associated with significant increase in median progression-free survival (PFS) of ovarian cancer patients receiving adjuvant platinum-taxane-based chemotherapy (P = 0.008). Multivariate analysis revealed that BRCA1 promoter methylation remains a favorable factor in regard to PFS (HR 0.52; 95 % CI 0.32-0.85, P = 0.009) after adjustment for other prognostic factors. Under the patients with recurrent disease, BRCA1 promoter methylation was associated with significant longer median PFS of 18.5 months in comparison with 12.8 months PFS for patients without BRCA1 promoter methylation.
BRCA1 promoter methylation is predictive for better response to platinum-taxane-based therapy in EOC.
Journal of Cancer Research and Clinical Oncology 05/2014; 140(9). DOI:10.1007/s00432-014-1704-5 · 3.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This chapter describes the continuing paramount importance of the histological diagnosis of Barrett's cancer and its prestages. It discusses the possible role of molecular predictors for estimation of the malignant potential of the prestages. The discussion includes a brief overview of the current experience with “biomarkers” for the diagnosis of Barrett's carcinogenesis, addressing the question of why their practical values have proven limited so far. The chapter also discusses the relationship between inflammation and Barrett's carcinogenesis, particularly for reflux esophagitis, including the role of genetic and epigenetic alterations in Barrett's carcinogenesis and the role of inflammation-associated reactive oxygen and nitrogen species (ROSs/RNSs).
Cancer and Inflammation Mechanisms, 04/2014: pages 213-222; , ISBN: 9781118160305
[Show abstract][Hide abstract] ABSTRACT: The production of hydrogen peroxide (H2 O2 ) drives tumourigenesis in ulcerative colitis (UC). Recently, we showed that H2 O2 activates DNA damage checkpoints in human colonic epithelial cells (HCEC) through c-Jun N-terminal Kinases (JNK) that induces p21(WAF1) . Moreover, caspases circumvented the G1/S and intra-S checkpoints, and cells accumulated in G2/M. The latter observation raised the question of whether repeated H2 O2 exposures alter JNK activation, thereby promoting a direct passage of cells from G2/M arrest to driven cell cycle progression. Here, we report that increased proliferation of repeatedly H2 O2 -exposed HCEC cells (C-cell cultures) was associated with (i) increased phospho-p46 JNK, (ii) decreased total JNK and phospho-p54 JNK and (iii) p21(WAF1) down-regulation. Altered JNK activation and p21(WAF1) down-regulation were accompanied by defects in maintaining G2/M and mitotic spindle checkpoints through adaptation, as well as by apoptosis resistance following H2 O2 exposure. This may cause increased proliferation of C-cell cultures, a defining initiating feature in the inflammation-carcinoma pathway in UC. We further suggest that dysregulated JNK activation is attributed to a non-apoptotic function of caspases, causing checkpoint adaptation in C-cell cultures. Additionally, loss of cell-contact inhibition and the overcoming of senescence, hallmarks of cancer, contributed to increased proliferation. Furthermore, there was evidence that p54 JNK inactivation is responsible for loss of cell-contact inhibition. We present a cellular model of UC and suggest a sinusoidal pattern of proliferation, which is triggered by H2 O2 -induced reactive oxygen species generation, involving an interplay between JNK activation/inactivation, p21(WAF1) , c-Fos, c-Jun/phospho-c-Jun, ATF2/phospho-ATF2, β-catenin/TCF4-signalling, c-Myc, CDK6 and Cyclin D2, leading to driven cell cycle progression.
Journal of Cellular and Molecular Medicine 10/2013; 17(12). DOI:10.1111/jcmm.12150 · 4.01 Impact Factor