Expression of GBGT1 is epigenetically regulated by DNA methylation in ovarian cancer cells

被引：12

作者：

Jaco, Francis ^{[1
,2
,3
]}

Hitchins, Megan P. ^{[4
]}

Fedier, Andre ^{[1
,2
]}

Brennan, Kevin ^{[4
]}

Nixdorf, Sheri ^{[3
]}

Hacker, Neville F. ^{[5
]}

Ward, Robyn ^{[3
]}

Heinzelmann-Schwarz, Viola A. ^{[1
,2
,3
,5
]}

机构：

[1] Univ Basel Hosp, Dept Biomed, Gynecol Res Grp, CH-4013 Basel, Switzerland

[2] Univ Basel, CH-4013 Basel, Switzerland

[3] Univ New S Wales, Prince Wales Clin Sch, Lowy Canc Res Ctr, Adult Canc Program, Sydney, NSW 2052, Australia

[4] Stanford Univ, Stanford Sch Med, Dept Med, Stanford, CA 94305 USA

[5] Royal Hosp Women, Gynaecol Canc Ctr, Sch Womens & Childrens Hlth, Sydney, NSW 2031, Australia

来源：

BMC MOLECULAR BIOLOGY | 2014年 / 15卷

基金：

瑞士国家科学基金会;

关键词：

Globo series; Glycosphingolipids; Forssman antigen; Epigenetics; DNA methylation; Transcriptional regulation; FORSSMAN GLYCOLIPIDS; ANTIGEN; ASSOCIATION; BREAST; HPA; 5-AZA-2'-DEOXYCYTIDINE; SYNTHETASE; GLOBOSIDE; GENE; TOOL;

D O I：

10.1186/1471-2199-15-24

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: The GBGT1 gene encodes the globoside alpha-1,3-N-acetylgalactosaminyltransferase 1. This enzyme catalyzes the last step in the multi-step biosynthesis of the Forssman (Fs) antigen, a pentaglycosyl ceramide of the globo series glycosphingolipids. While differential GBGT1 mRNA expression has been observed in a variety of human tissues being highest in placenta and ovary, the expression of GBGT1 and the genes encoding the glycosyltransferases and glycosidases involved in the biosynthesis of Fs as well as the possible involvement of DNA methylation in transcriptional regulation of GBGT1 expression have not yet been investigated. Results: RT-qPCR profiling showed high GBGT1 expression in normal ovary surface epithelial (HOSE) cell lines and low GBGT1 expression in all (e.g. A2780, SKOV3) except one (OVCAR3) investigated ovarian cancer cell lines, a finding that was confirmed by Western blot analysis. Hierarchical cluster analysis showed that GBGT1 was even the most variably expressed gene of Fs biosynthesis-relevant glycogenes and among the investigated cell lines, whereas NAGA which encodes the alpha-N-acetylgalactosaminidase hydrolyzing Fs was not differentially expressed. Bisulfite- and COBRA-analysis of the CpG island methylation status in the GBGT1 promoter region demonstrated high or intermediate levels of GBGT1 DNA methylation in all ovarian cancer cell lines (except for OVCAR3) but marginal levels of DNA methylation in the two HOSE cell lines. The extent of DNA methylation inversely correlated with GBGT1 mRNA and protein expression. Bioinformatic analysis of GBGT1 in The Cancer Genome Atlas ovarian cancer dataset demonstrated that this inverse correlation was also found in primary ovarian cancer tissue samples confirming our cell line-based findings. Restoration of GBGT1 mRNA and protein expression in low GBGT1-expressing A2780 cells was achieved by 5-aza-2'-deoxycytidine treatment and these treated cells exhibited increased helix pomatia agglutinin-staining, reflecting the elevated presence of Fs disaccharide on these cells. Conclusions: GBGT1 expression is epigenetically silenced through promoter hypermethylation in ovarian cancer. Our findings not only suggest an involvement of DNA methylation in the synthesis of Fs antigen but may also explain earlier studies showing differential GBGT1 expression in various human tissue samples and disease stages.

引用

页数：13

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