title: Epigenetic inactivation of heparan sulfate (glucosamine) 3-O-sulfotransferase 2 in lung cancer and its role in tumorigenesis.
abstract: BACKGROUND
  This study was aimed at investigating the functional significance of heparan sulfate (glucosamine) 3-O-sulfotransferase 2 (HS3ST2) hypermethylation in non-small cell lung cancer (NSCLC).
  
  
  RESULTS
  HS3ST2 hypermethylation was characterized in six lung cancer cell lines, and its clinical significance was analyzed using 298 formalin-fixed paraffin-embedded tissues and 26 fresh-frozen tissues from 324 NSCLC patients. MS-HRM (methylation-specific high-resolution melting) and EpiTYPER(TM) assays showed substantial hypermethylation of CpG island at the promoter region of HS3ST2 in six lung cancer cell lines. The silenced gene was demethylated and re-expressed by treatment with 5-aza-2'-deoxycytidine (5-Aza-dC). A promoter assay also showed the core promoter activity of HS3ST2 was regulated by methylation. Exogenous expression of HS3ST2 in lung cancer cells H460 and H23 inhibited cell migration, invasion, cell proliferation and whereas knockdown of HS3ST2 in NHBE cells induced cell migration, invasion, and cell proliferation in vitro. A negative correlation was observed between mRNA and methylation levels of HS3ST2 in 26 fresh-frozen tumors tissues (ρ = -0.51, P = 0.009; Spearman's rank correlation). HS3ST2 hypermethylation was found in 95 (32%) of 298 primary NSCLCs. Patients with HS3ST2 hypermethylation in 193 node-negative stage I-II NSCLCs with a median follow-up period of 5.8 years had poor overall survival (hazard ratio = 2.12, 95% confidence interval = 1.25-3.58, P = 0.005) compared to those without HS3ST2 hypermethylation, after adjusting for age, sex, tumor size, adjuvant therapy, recurrence, and differentiation.
  
  
  CONCLUSIONS
  The present study suggests that HS3ST2 hypermethylation may be an independent prognostic indicator for overall survival in node-negative stage I-II NSCLC.
authors.affiliation: Cancer Genomics Branch, Research Institute, National Cancer Center, Goyang-si, Korea.
authors.forename: Jung-Ah
authors.initials: JA
authors.lastname: Hwang
authors.forename: Yujin
authors.initials: Y
authors.lastname: Kim
authors.forename: Seung-Hyun
authors.initials: SH
authors.lastname: Hong
authors.forename: Jieun
authors.initials: J
authors.lastname: Lee
authors.forename: Yong Gu
authors.initials: YG
authors.lastname: Cho
authors.forename: Ji-Youn
authors.initials: JY
authors.lastname: Han
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Kim
authors.forename: Joungho
authors.initials: J
authors.lastname: Han
authors.forename: Young Mog
authors.initials: YM
authors.lastname: Shim
authors.forename: Yeon-Su
authors.initials: YS
authors.lastname: Lee
authors.forename: Duk-Hwan
authors.initials: DH
authors.lastname: Kim
mesh_terms: D000328:Adult; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D063646:Carcinogenesis / Q000187:drug effects / Q000235:genetics*; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics; D000077209:Decitabine; D005260:Female; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics* / Q000473:pathology*; D008297:Male; D009361:Neoplasm Invasiveness; D010641:Phenotype; D011379:Prognosis; D011401:Promoter Regions, Genetic / Q000235:genetics; D016016:Proportional Hazards Models; D015238:Sulfotransferases / Q000172:deficiency* / Q000235:genetics*
chemical_list: D000077209:Decitabine; D015238:Sulfotransferases; C093554:heparin-glucosamine 3-O-sulfotransferase; D001374:Azacitidine