Analysis of different methylation patterns of histone H3 between andro-gen-dependent and androgen-independent prostate cancer cells by heavy methyl SILAC
10.3969/j.issn.1000-4718.2014.09.010
- VernacularTitle:重甲基 SILAC 技术分析雄激素依赖性和非依赖性前列腺癌细胞组蛋白 H3甲基化的差异
- Author:
Qilin WANG
;
Ruiqian LI
;
Congguo JIN
;
Bin ZHAO
;
Guoying ZHANG
;
Yu BAI
;
Yonghong LEI
- Publication Type:Journal Article
- Keywords:
Prostatic neoplasms;
Histone H3;
Methylation
- From:
Chinese Journal of Pathophysiology
2014;(9):1595-1602
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To study the epigenetic mechanisms involved in the evolution of prostate cancer from an an-drogen-dependent state to an androgen-independent state , and the global difference of histone H 3 methylation between an-drogen-dependent and -independent prostate cancer cells .METHODS:The methylation sites and patterns of histone H 3 in androgen-dependent prostate cancer cell line LNCaP and androgen-independent prostate cancer cell line DU 145 were ana-lyzed by heavy methyl stable isotope labeling with amino acids in cell culture ( SILAC) coupled with 2D LC-MS/MS.West-ern blotting was used to verify the results from MS .The differential expression of related methylases and demethylases was tested by real-time PCR.RESULTS:Five methylation sites on histone H3 were found in both cell lines, the patterns of which were as follows: H3K14me2, H3R17me1, H3K36me1, H3K36me2, H3K36me3, H3R72me2, H3K79me1 and H3K79me2.There were 2 different peptides both containing methylated H 3K36,“KSAPATGGVKKPHR” and“KSAPSTG-GVKKPHR”, which were different from the 31th amino acid residue “A” and “S”.The former peptide belonging to his-tone H3 variants, H31T, H31 and H32, was mainly identified in DU145 cells, the total peptide counts of which were much more than that of the latter peptide belonging to histone H 3 variant H31T, suggesting that these 2 cell lines expressed differ-ent histone H3 variants.Mono-and dimethylation of H3K36 were not different between these 2 cell lines, but the trimethyl-ation was significantly higher in DU 145 cells than that in LNCaP cells .Many H3K36 demethyltransferases were decreased in DU145 cells compared with LNCaP cells .CONCLUSION: The differential expression of histone H 3 variants and H3K36 demethyltransferases may result in up-regulation of H3K36 tri-methylation during the evolution of prostate cancer from an androgen-dependent state to an androgen-independent state .
