title: Methylation-associated silencing of heparan sulfate D-glucosaminyl 3-O-sulfotransferase-2 (3-OST-2) in human breast, colon, lung and pancreatic cancers.
abstract: Aberrant CpG methylations play important roles in cancer development and progression. In this study, aberrant methylations in human breast cancer were searched for using methylation-sensitive representational difference analysis (MS-RDA). A CpG island (CGI) in the 5' region of the heparan sulfate D-glucosaminyl 3-O-sulfotransferase-2 (3-OST-2) gene was found to be hypermethylated, while its exon 2 was hypomethylated. In seven breast cancer cell lines, hypermethylation of the 5' region and loss of 3-OST-2 expression were observed. Treatment with a demethylating agent, 5-aza-2'-deoxycytidine, removed the methylation of the CGI in the 5' region and restored its expression, demonstrating silencing of the 3-OST-2 gene. Methylation-specific PCR (MSP) analysis in 85 primary breast cancers showed that the hypermethylation of the CGI in the 5' region was present in 75 (88%) of them. Quantitative reverse transcriptase-PCR (RT-PCR) analysis in 37 primary breast cancers showed that the average expression level was decreased in them. Further, MSP analysis in primary colon, lung and pancreatic cancers showed that hypermethylation of the CGI in the 5' region was present in the colon (8/10, 80%), lung (7/10, 70%) and pancreatic (10/10, 100%) cancers. These results showed that silencing of 3-OST-2 was present in a wide range of human cancers. The 3-OST-2 gene encodes an enzyme involved in the final modification step of heparan sulfate proteoglycans (HSPGs), and its silencing is expected to result in abnormal modification of HSPGs and abnormal signal transduction. From the high incidence, silencing of the 3-OST-2 gene is expected to have high diagnostic, and potentially therapeutic, values.
authors.affiliation: National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Asada
authors.forename: Takashi
authors.initials: T
authors.lastname: Fukutomi
authors.forename: Eriko
authors.initials: E
authors.lastname: Okochi
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yagi
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hasegawa
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Asahara
authors.forename: Takashi
authors.initials: T
authors.lastname: Sugimura
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Ushijima
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D003110:Colonic Neoplasms / Q000235:genetics*; D018899:CpG Islands; D019175:DNA Methylation; D005091:Exons; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods; D010190:Pancreatic Neoplasms / Q000235:genetics*; D015238:Sulfotransferases / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D015238:Sulfotransferases; C102180:heparan sulfate D-glucosaminyl 3-O-sulfotransferase