Overexpression of Mucin 13 due to Promoter Methylation Promotes Aggressive Behavior in Ovarian Cancer Cells
2
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
Purpose: Recent discoveries suggest that aberrant DNA methylation provides cancercells with advanced metastatic properties. However, the precise regulatory mechanismscontrolling metastasis genes and their role in metastatic transformation are largely unknown. To address epigenetically-regulated gene products involved in ovarian cancer metastasis, we examined the mechanisms regulating mucin 13 (MUC13) expression and its influence on aggressive behaviors of ovarian malignancies. Materials and Methods: We injected SK-OV-3 ovarian cancer cells peritoneallyinto nude mice to mimic human ovarian tumor metastasis. Overexpression of MUC13 mRNA was detected in metastatic implants from the xenografts by expressionmicroarray analysis and quantitative reverse-transcription polymerase chain reaction(qRT-PCR). The DNA methylation status within the MUC13 promoter region was determined using bisulfite sequencing PCR and quantitative methylation-specificPCR. We evaluated the effects of exogenous MUC13 on cell invasion and migrationusing in vitro transwell assays. Results: MUC13 mRNA expression was up-regulated,and methylation of specific CpG sites within the promoter was reduced in the metastatic implants relative to those in wild-type SK-OV-3 cells. Addition of a DNA methyltransferase inhibitor to SK-OV-3 cells induced MUC13 expression, thereby implying epigenetic regulation of MUC13 by promoter methylation. MUC13 overexpressionincreased migration and invasiveness, compared to control cells, suggestingaberrant up-regulation of MUC13 is strongly associated with progression of aggressivebehaviors in ovarian cancer. Conclusion: We provide novel evidence for epigenetic regulation of MUC13 in ovarian cancer. We suggest that the DNA methylationstatus within the MUC13 promoter region may be a potential biomarker of aggressive behavior in ovarian cancer.
Abstract
Purpose: Recent discoveries suggest that aberrant DNA methylation provides cancercells with advanced metastatic properties. However, the precise regulatory mechanismscontrolling metastasis genes and their role in metastatic transformation are largely unknown. To address epigenetically-regulated gene products involved in ovarian cancer metastasis, we examined the mechanisms regulating mucin 13 (MUC13) expression and its influence on aggressive behaviors of ovarian malignancies. Materials and Methods: We injected SK-OV-3 ovarian cancer cells peritoneallyinto nude mice to mimic human ovarian tumor metastasis. Overexpression of MUC13 mRNA was detected in metastatic implants from the xenografts by expressionmicroarray analysis and quantitative reverse-transcription polymerase chain reaction(qRT-PCR). The DNA methylation status within the MUC13 promoter region was determined using bisulfite sequencing PCR and quantitative methylation-specificPCR. We evaluated the effects of exogenous MUC13 on cell invasion and migrationusing in vitro transwell assays. Results: MUC13 mRNA expression was up-regulated,and methylation of specific CpG sites within the promoter was reduced in the metastatic implants relative to those in wild-type SK-OV-3 cells. Addition of a DNA methyltransferase inhibitor to SK-OV-3 cells induced MUC13 expression, thereby implying epigenetic regulation of MUC13 by promoter methylation. MUC13 overexpressionincreased migration and invasiveness, compared to control cells, suggestingaberrant up-regulation of MUC13 is strongly associated with progression of aggressivebehaviors in ovarian cancer. Conclusion: We provide novel evidence for epigenetic regulation of MUC13 in ovarian cancer. We suggest that the DNA methylationstatus within the MUC13 promoter region may be a potential biomarker of aggressive behavior in ovarian cancer.
참고문헌 (22)
- Lengyel E, "Ovarian cancer development and metastasis", Am J Pathol, 177, pp.1053~1064 (2010)
- Kipps E, "Meeting the challenge of ascites in ovarian cancer: new avenues for therapy and research", Nat Rev Cancer, 13, pp.273~282 (2013)
- Rose MC, "Respiratory tract mucin genes and mucin glycoproteins in health and disease", Physiol Rev, 86, pp.245~278 (2006)
- Singh AP, "Clinical potential of mucins in diagnosis, prognosis, and therapy of ovarian cancer", Lancet Oncol, 9, pp.1076~1085 (2008)
- Hollingsworth MA, "Mucins in cancer: protection and control of the cell surface", Nat Rev Cancer, 4, pp.45~60 (2004)
- Gupta BK, "Increased expression and aberrant localization of mucin 13 in metastatic colon cancer", J Histochem Cytochem, 60, pp.822~831 (2012)
- Ringel J, "The MUC gene family: their role in diagnosis and early detection of pancreatic cancer", Mol Cancer, 2, pp.9~ (2003)
- Brockhausen I, "Mechanisms underlying aberrant glycosylation of MUC1 mucin in breast cancer cells", Eur J Biochem, 233, pp.607~617 (1995)
- Rakha EA, "Expression of mucins (MUC1, MUC2, MUC3, MUC4, MUC5AC and MUC6) and their prognostic significance in human breast cancer", Mod Pathol, 18, pp.1295~1304 (2005)
- Rosen DG, "Potential markers that complement expression of CA125 in epithelial ovarian cancer", Gynecol Oncol, 99, pp.267~277 (2005)
- Dong Y, "Expression of MUC1 and MUC2 mucins in epithelial ovarian tumours", J Pathol, 183, pp.311~317 (1997)
- Giuntoli RL 2nd, "Mucin gene expression in ovarian cancers", Cancer Res, 58, pp.5546~5550 (1998)
- Chauhan SC, "Expression and functions of transmembrane mucin MUC13 in ovarian cancer", Cancer Res, 69, pp.765~774 (2009)
- Canney PA, "Ovarian cancer antigen CA125: a prospective clinical assessment of its role as a tumour marker", Br J Cancer, 50, pp.765~769 (1984)
- Yin BW, "Ovarian cancer antigen CA125 is encoded by the MUC16 mucin gene", Int J Cancer, 98, pp.737~740 (2002)
- Budiu RA, "Soluble MUC1 and serum MUC1-specific antibodies are potential prognostic biomarkers for platinum-resistant ovarian cancer", Cancer Immunol Immunother, 60, pp.975~984 (2011)
- Chauhan SC, "Aberrant expression of MUC4 in ovarian carcinoma: diagnostic significance alone and in combination with MUC1 and MUC16 (CA125)", Mod Pathol, 19, pp.1386~1394 (2006)
- Williams SJ, "Muc13, a novel human cell surface mucin expressed by epithelial and hemopoietic cells", J Biol Chem, 276, pp.18327~18336 (2001)
- Maher DM, "Mucin 13: structure, function, and potential roles in cancer pathogenesis", Mol Cancer Res, 9, pp.531~537 (2011)
- Walsh MD, "The MUC13 cell surface mucin is highly expressed by human colorectal carcinomas", Hum Pathol, 38, pp.883~892 (2007)
- Shimamura T, "Overexpression of MUC13 is associated with intestinal-type gastric cancer", Cancer Sci, 96, pp.265~273 (2005)
- Chauhan SC, "MUC13 mucin augments pancreatic tumorigenesis", Mol Cancer Ther, 11, pp.24~33 (2012)