Journal of Andrology
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Published-Ahead-of-Print October 16, 2008, DOI:10.2164/jandrol.108.005785

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Homo Sapiens Lactate Dehydrogenase c (Ldhc) Gene Expression in Cancer Cells is Regulated by Transcription Factor Sp1, CREB and CpG Island Methylation

HuangHui Tang * and Erwin Goldberg

* To whom correspondence should be addressed. E-mail: h-tang2{at}northwestern.edu.

The human testis-specific lactate dehydrogenase c gene (hLdhc) is transcribed only in cells of the germinal epithelium. Recently hLdhc was reported to express in a broad spectrum of tumors with relatively high frequency in lung cancer, melanoma and breast cancer, and in some prostate cancers. Two melanoma cell lines, A375M and C81-61, were identified that express the hLdhc gene and were used to characterize the hLdhc promoter and explore transcriptional regulation of this gene. A 110-bp core promoter, including a conserved GC-box and cAMP-responsive element (CRE), were identified as essential for basal promoter activity. The methylation status of the CpG island (CGI) in the hLdhc core promoter sequence was analyzed in hLdhc expressing and non-expressing cells and human prostate tumor tissues. The CGI in two cell lines expressing the gene were hypomethylated while the DNAs from cells that did not express hLdhc were hypermethylated. The role of methylation in regulating this promoter was confirmed by experimental induction of hLdhc transcription with the methylation inhibitor 5'Aza-deoxycytidine. Quantative analyses of the methylation level in the CpG island were performed in prostate tumor tissues by pyrosequencing. Overall, these experiments demonstrated that hLdhc expression in cancer cells was regulated by transcription factor Sp1 and CREB and promoter CpG island methylation. In addition these findings suggest the possibility of developing a biomarker for cancer diagnosis/prognosis based on DNA methylation of the Ldhc gene.


Key words: CpG island • caner/testis antigen • ldhc • methylation • promoter




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