transforming-growth-factor-beta and Adenocarcinoma--Clear-Cell

Topics: Adenocarcinoma, Clear Cell; Cell Division; Endometriosis; Female; Humans; Ovarian Neoplasms; Prognosis; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor beta

Expression of Nodal, a member of the TGF-β superfamily, is commonly absent in differentiated tissues, while its re-expression occurs in a variety of human malignancy. However, little is known about its involvement in ovarian tumorigenesis. Herein, we focused on the functional roles of Nodal in ovarian endometriosis-carcinoma lesions.. Regulation and function of Nodal and its associated molecules, including Smad2, GSK-3β, and several cell kinetics-related molecules, were assessed using clinical samples consisting of 108 ovarian carcinomas and 33 endometriotic lesions, as well as ES-2 (ovarian clear cell carcinoma; OCCCa) and Ishikawa (endometrial carcinoma) cell lines.. Nodal expression was significantly higher in endometriosis and OCCCa lesions as compared to that of non-OCCCas, with positive correlations to phosphorylated forms of both Smad2 (pSmad2) and GSK-3β. When compared to endometriotic lesions, the expression of Nodal and pSmad2 was significantly decreased in OCCCa. Treatment of Ishikawa cells with TGF-β1 resulted in transcriptional upregulation of Nodal, along with increased pSmad2 expression, while inhibition of GSK-3β also induced an increase in Nodal expression at the posttranslational level. Both ES-2 and Ishikawa cells stably overexpressing Nodal had increased susceptibility to apoptosis in response to treatment with cisplatin and doxorubicin, respectively, together with higher cleaved caspase-3 expression and decreased Bcl2/Bax ratio. Moreover, the stable Nodal-overexpressing cells showed reduced cell proliferation, along with increased expression of p27. These findings suggest that Nodal is a multifunctional cytokine involved in the modulation of cell kinetics in ovarian endometriosis-OCCCa lesions.

Topics: Adenocarcinoma, Clear Cell; Adult; Aged; Aged, 80 and over; Apoptosis; Cell Proliferation; Endometriosis; Female; Humans; Middle Aged; Nodal Protein; Ovarian Neoplasms; Phosphorylation; Signal Transduction; Smad2 Protein; Transforming Growth Factor beta; Up-Regulation

Topics: Adenocarcinoma, Clear Cell; Adult; Aged; Aged, 80 and over; Epithelial-Mesenchymal Transition; Female; Glycogen Synthase Kinase 3 beta; Humans; Left-Right Determination Factors; Middle Aged; Neoplastic Stem Cells; Ovarian Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; Snail Family Transcription Factors; Transforming Growth Factor beta

Using pharmacologic unmasking and genome-wide differential methylation analysis, we identified a novel methylated gene in ovarian cancers.. Two ovarian cancer cells (OVCAR-3, ES-2) that showed synergistic growth inhibition by 5-aza-dC and cisplatin were selected. After treatment with 5-aza-dC, differential expression profiles were compared using microarray that contained 38,500 genes. Reactivation of candidate genes and their promoter methylation were validated by real-time RT-PCR, MS-PCR and bisulfite sequencing. Methylation status was tested by MS-PCR in 56 patients with epithelial ovarian cancer and compared to the 38 normal ovarian tissues.. We identified 103 candidate genes that were reactivated by 5-aza-dC treatment. Among those, SFN and TGFBI were commonly reactivated in both cells. Since SFN is a well known methylated marker, we selected TGFBI for further validation. Bisulfite sequencing revealed complete promoter methylation in ES-2 and partial methylation in OVCAR-3. In addition, silencing of TGFBI at the transcription level was reversed by 5-aza-dC treatment. TGFBI methylation was observed in 23 out of 38 (60.5%) cases of ovarian cancer, in no normal ovarian tissues (0 of 38, P=0.001), and in 5 out of 18 (27.8%) borderline tumors (P=0.044). In our cohort, we did not observe any association between methylation of TGFBI and clinicopathologic variables or clinical outcomes.. Our results confirm that TGFBI is frequently methylated in ovarian cancer. Its methylation can be used as a novel epigenetic biomarker in discriminating ovarian cancer from non-cancer or borderline tumors.

Topics: Adenocarcinoma; Adenocarcinoma, Clear Cell; Adult; Aged; Cell Line, Tumor; DNA Methylation; Epithelial Cells; Extracellular Matrix Proteins; Female; Humans; Middle Aged; Oligonucleotide Array Sequence Analysis; Ovarian Neoplasms; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta

Transforming growth factor-beta1 (TGF-beta1) has a biphasic effect on the growth of renal epithelial cells. In transformed cells, TGF-beta1 appears to accelerate the proliferation of malignant cells. The diverse cellular functions of TGF-beta1 are regulated by three high-affinity serine/threonine kinase receptors, namely TbetaRI, TbetaRII and TbetaRIII. The renal serine protease tissue kallikrein acts on its endogenous protein substrate kininogen to form kinin peptides. The cellular actions of kinins are mediated through B1 and B2 G protein-coupled rhodopsin receptors. Both kinin peptides and TGF-beta1 are mitogenic, and therefore may play an important role in carcinogenesis. Experiments were designed to immunolabel tissue kallikrein, TGF-beta1, TbetaRII, TbetaRIII and kinin receptors using specific antibodies on serial sections of normal kidney and clear-cell renal carcinoma (CCRC) tissue, which included both the tumour and the adjacent renal parenchyma. The essential result was the localisation of tissue kallikrein, kinin B 1 and B 2 receptors and TGF-beta1 primarily on the cell membranes of CCRC cells. In the distal and proximal tubules of the renal parenchyma adjacent to the carcinoma (RPTAC), immunolabelling for tissue kallikrein was reduced, but the expression of kinin B1 and B2 receptors was enhanced. Immunolabelling for TbetaRII and TbetaRIII was more pronounced in the proximal tubules of the tissue adjacent to the carcinoma when compared to the normal kidney. The expression of tissue kallikrein, kinin receptors, and TbetaRII and TbetaRIII may be relevant to the parenchymal invasion and metastasis of clear-cell renal carcinoma.

Topics: Adenocarcinoma, Clear Cell; Humans; Kidney Neoplasms; Receptors, Bradykinin; Receptors, Transforming Growth Factor beta; Tissue Kallikreins; Transforming Growth Factor beta; Transforming Growth Factor beta1

The von Hippel-Lindau (VHL) tumor suppressor gene is mutated in patients with VHL disease and in the majority of patients with sporadic clear cell renal carcinoma (RCC). Overexpression of transforming growth factor (TGF) beta1 has been observed in patients with several cancers, including RCCs, with serum and urine levels correlating inversely with prognosis. We have demonstrated that the VHL tumor suppressor gene product represses TGF-beta1 mRNA and protein levels (approximately 3-4-fold) in 786-O RCC cells by decreasing the TGF-beta1 mRNA half-life. Exogenously added TGF-beta1 did not suppress the growth of 786-O cells in vitro, nor did the addition of neutralizing antibody (Ab) against TGF-beta have any effect. Indeed, 786-O cells were found to express no TGF-beta type II receptor protein, thus allowing them to escape from the negative growth control of TGF-beta1. In contrast to the in vitro data, neutralizing Ab to TGF-beta inhibited tumorigenesis and, in some cases, regressed established 786-O tumors in athymic mice. Immunohistochemistry for von Willebrand's factor revealed a 3-4-fold lower tumor microvessel count in the mice treated with TGF-beta Ab compared to controls, suggesting that the Ab was inhibiting angiogenesis. Our findings indicate that TGF-beta1 is a novel target for the VHL tumor suppressor and that antagonizing its paracrine action may provide novel avenues for treatment of RCCs as well as other tumors that secrete TGF-beta1.

Topics: Adenocarcinoma, Clear Cell; Animals; Antibodies, Monoclonal; Gene Deletion; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Half-Life; Humans; Kidney Neoplasms; Ligases; Mice; Mice, Mutant Strains; Mice, Nude; Neoplasm Proteins; Neoplasm Transplantation; Neovascularization, Pathologic; Proteins; Recombinant Fusion Proteins; RNA Processing, Post-Transcriptional; RNA, Messenger; RNA, Neoplasm; Transfection; Transforming Growth Factor beta; Transplantation, Heterologous; Tumor Cells, Cultured; Tumor Suppressor Proteins; Ubiquitin-Protein Ligases; Von Hippel-Lindau Tumor Suppressor Protein; von Willebrand Factor