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).

METHODOLOGY/ PRINCIPAL FINDINGS:

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/ SIGNIFICANCE:

The present study suggests that HS3ST2 hypermethylation may be an independent prognostic indicator for overall survival in node-negative stage I-II NSCLC.

• Authors: Jung-Ah Hwang, Yujin Kim, Seung-Hyun Hong, Jieun Lee, Yong Gu Cho, Ji-Youn Han, Young-Ho Kim, Joungho Han, Young Mog Shim, Yeon-Su Lee, Duk-Hwan Kim
• Journal: PloS one (PLoS One) Vol. 8 Issue 11 Pg. e79634 ( 2013) ISSN: 1932-6203 [Electronic] United States
• PMID: 24265783 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Decitabine
• Sulfotransferases
• heparin-glucosamine 3-O-sulfotransferase
• Azacitidine

Topics

• Adult
• Azacitidine (analogs & derivatives, pharmacology)
• Carcinogenesis (drug effects, genetics)
• Carcinoma, Non-Small-Cell Lung (diagnosis, enzymology, genetics, pathology)
• Cell Line, Tumor
• Cell Movement (drug effects, genetics)
• Cell Proliferation (drug effects)
• DNA Methylation (drug effects, genetics)
• Decitabine
• Female
• Gene Knockdown Techniques
• Gene Silencing
• Humans
• Lung Neoplasms (diagnosis, enzymology, genetics, pathology)
• Male
• Neoplasm Invasiveness
• Phenotype
• Prognosis
• Promoter Regions, Genetic (genetics)
• Proportional Hazards Models
• Sulfotransferases (deficiency, genetics)