Regulation of MUC6 Methylation Correlates with Progression of Gastric Cancer
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: This study aimed to investigate the mechanistic downregulation of mucin 6 (MUC6) and its influence on the progression of gastric cancer (GC). Materials and Methods: The expression of MUC6 was examined in 40 GC patients. The methylation status of the MUC6 promoter region was investigated using GC cell lines and GC tissue specimens by immunohistochemistry and/or quantitative polymerase chain reaction (qPCR). MUC6 was knocked down in the gastric epithelial cells (GES-1) cell and overexpressed in the SGC7901 cell. The effects of MUC6 knockdown and overexpression on cell migration and invasion were examined using Transwell assays. The effects of demethylation and methylation on MUC6 expression were examined by western blot, qPCR, or double luciferase reporter assays. Results: The expression of MUC6 in GC with lymph node metastasis and poor pathological stage was significantly lower than that in GC without lymph node metastasis and good pathological stage, respectively. While cell migration and invasion were significantly decreased after overexpression of MUC6, these abilities significantly increased after the knockdown of MUC6. The methylation levels of MUC6 in GC tissues and GC cell lines were significantly higher than those in para-cancerous tissues and normal GES. Promoter methylation could significantly reduce the binding of related transcription factors to the MUC6 promoter. The expression of MUC6 increased with the concentration and time of action of demethylation drugs. Conclusion: Expression of MUC6 was regulated by promotor methylation. This methylation of the MUC6 promoter may lead to significant downregulation of MUC6 in GC and promote the progression of GC.
Abstract
Purpose: This study aimed to investigate the mechanistic downregulation of mucin 6 (MUC6) and its influence on the progression of gastric cancer (GC). Materials and Methods: The expression of MUC6 was examined in 40 GC patients. The methylation status of the MUC6 promoter region was investigated using GC cell lines and GC tissue specimens by immunohistochemistry and/or quantitative polymerase chain reaction (qPCR). MUC6 was knocked down in the gastric epithelial cells (GES-1) cell and overexpressed in the SGC7901 cell. The effects of MUC6 knockdown and overexpression on cell migration and invasion were examined using Transwell assays. The effects of demethylation and methylation on MUC6 expression were examined by western blot, qPCR, or double luciferase reporter assays. Results: The expression of MUC6 in GC with lymph node metastasis and poor pathological stage was significantly lower than that in GC without lymph node metastasis and good pathological stage, respectively. While cell migration and invasion were significantly decreased after overexpression of MUC6, these abilities significantly increased after the knockdown of MUC6. The methylation levels of MUC6 in GC tissues and GC cell lines were significantly higher than those in para-cancerous tissues and normal GES. Promoter methylation could significantly reduce the binding of related transcription factors to the MUC6 promoter. The expression of MUC6 increased with the concentration and time of action of demethylation drugs. Conclusion: Expression of MUC6 was regulated by promotor methylation. This methylation of the MUC6 promoter may lead to significant downregulation of MUC6 in GC and promote the progression of GC.
참고문헌 (28)
- Pinto-de-Sousa J, "MUC5B expression in gastric carcinoma : relationship with clinico-pathological parameters and with expression of mucins MUC1, MUC2, MUC5AC and MUC6", Virchows Arch, 444, pp.224~230 (2004)
- Senapati S, "The MUC gene family : their role in the diagnosis and prognosis of gastric cancer", Histol Histopathol, 23, pp.1541~1552 (2008)
- Jia Y, "A comprehensive analysis of common genetic variation in MUC1, MUC5AC, MUC6 genes and risk of stomach cancer", Cancer Causes Control, 21, pp.313~321 (2010)
- Nakayama J, "Dual roles of gastric gland mucin-specific O-glycans in prevention of gastric cancer", Acta Histochem Cytochem, 47, pp.1~9 (2014)
- Babu SD, "Expression profile of mucins(MUC2, MUC5AC and MUC6)in Helicobacter pylori infected pre-neoplastic and neoplastic human gastric epithelium", Mol Cancer, 5, pp.10~ (2006)
- Park KK, "Correlations of human epithelial growth factor receptor 2 overexpression with MUC2, MUC5AC, MUC6, p53, and clinicopathological characteristics in gastric cancer patients with curative resection", Gastroenterol Res Pract, 2015, pp.946359~ (2015)
- Reis CA, "Intestinal metaplasia of human stomach displays distinct patterns of mucin (MUC1, MUC2, MUC5AC, and MUC6) expression", Cancer Res, 59, pp.1003~1007 (1999)
- Reis CA, "Immunohistochemical study of the expression of MUC6 mucin and co-expression of other secreted mucins(MUC5AC and MUC2)in human gastric carcinomas", J Histochem Cytochem, 48, pp.377~388 (2000)
- Ho SB, "Mucin gene expression in normal, preneoplastic, and neoplastic human gastric epithelium", Cancer Res, 55, pp.2681~2690 (1995)
- Zheng H, "MUC6 down-regulation correlates with gastric carcinoma progression and a poor prognosis : an immunohistochemical study with tissue microarrays", J Cancer Res Clin Oncol, 132, pp.817~823 (2006)
- Kwon JA, "Short rare MUC6 minisatellites-5 alleles influence susceptibility to gastric carcinoma by regulating gene", Hum Mutat, 31, pp.942~949 (2010)
- Terada T, "An immunohistochemical study of primary signet-ring cell carcinoma of the stomach and colorectum: II. Expression of MUC1, MUC2, MUC5AC, and MUC6 in normal mucosa and in 42cases", Int J Clin Exp Pathol, 6, pp.613~621 (2013)
- Shiratsu K, "Loss of gastric gland mucinspecific O-glycan is associated with progression of differentiatedtype adenocarcinoma of the stomach", Cancer Sci, 105, pp.126~133 (2014)
- Pigny P, "Human mucin genes assigned to 11p15.5: identification and organization of a cluster of genes", Genomics, 38, pp.340~352 (1996)
- Vincent A, "Epigenetic regulation(DNA methylation, histone modifications)of the 11p15 mucin genes(MUC2, MUC5AC, MUC5B, MUC6)in epithelial cancer cells", Oncogene, 26, pp.6566~6576 (2007)
- Gratchev A, "Regulation of mucin MUC2 gene expression during colon carcinogenesis", Ann N Y Acad Sci, 859, pp.180~183 (1998)
- Siedow A, "De novo expression of the Muc2 gene in pancreas carcinoma cells is triggered by promoter demethylation", Tumour Biol, 23, pp.54~60 (2002)
- Mesquita P, "Role of site-specific promoter hypomethylation in aberrant MUC2 mucin expression in mucinous gastric carcinomas", Cancer Lett, 189, pp.129~136 (2003)
- Bartman AE, "The MUC6 secretory mucin gene is expressed in a wide variety of epithelial tissues", J Pathol, 186, pp.398~405 (1998)
- Garcia E, "MUC6 gene polymorphism in healthy individuals and in gastric cancer patients from northern Portugal", Cancer Epidemiol Biomarkers Prev, 6, pp.1071~1074 (1997)
- Williams SJ, "Two novel mucin genes down-regulated in colorectal cancer identified by differential display", Cancer Res, 59, pp.4083~4089 (1999)
- Perrais M, "Aberrant expression of human mucin gene MUC5B in gastric carcinoma and cancer cells. Identification and regulation of a distal promoter", J Biol Chem, 276, pp.15386~15396 (2001)
- Shi D, "The changes in MUC5AC expression in gastric cancer before and after Helicobacter pylori eradication", Clin Res Hepatol Gastroenterol, 38, pp.235~240 (2014)
- Jones PA, "The epigenomics of cancer", Cell, 128, pp.683~692 (2007)
- Toyota M, "Epigenetic changes in solid and hematopoietic tumors", Semin Oncol, 32, pp.521~530 (2005)
- Ushijima T, "Detection and interpretation of altered methylation patterns in cancer cells", Nat Rev Cancer, 5, pp.223~231 (2005)
- Bird AP, "Methylation-induced repression--belts, braces, and chromatin", Cell, 99, pp.451~454 (1999)
- Jones PA, "Cancer epigenetics comes of age", Nat Genet, 21, pp.163~167 (1999)