肝细胞癌中<i>ChREBP</i>基因CpG岛甲基化与mRNA表达的相关性

冉凤鸣; 罗成刚; 邱雪平; 黄一芳; 郑芳; 魏少忠

doi:10.3971/j.issn.1000-8578.2016.08.009

肝细胞癌中ChREBP基因CpG岛甲基化与mRNA表达的相关性

Correlation of ChREBP mRNA Expression and Its CpG Island Methylation in Human Hepatocellular Carcinoma

摘要

摘要:
目的检测肝细胞癌中ChREBP基因CpG岛甲基化水平及mRNA表达水平，并探讨两者的相关性。
方法收集肝细胞癌及癌旁冰冻组织90例，采用亚硫酸氢钠处理结合克隆测序检测ChREBP基因CpG岛内29个CpG位点的甲基化水平；采用荧光定量PCR检测其中31对新鲜冰冻组织中ChREBP基因mRNA表达水平，分析甲基化水平与mRNA表达之间的关系。
结果 ChREBP基因的5、6、7、14号CpG位点甲基化水平在肝细胞癌中显著低于其配对的癌旁组织（P<0.05）。其余CpG位点及整体甲基化在肝细胞癌与癌旁组织中差异无统计学意义（均P<0.05）。15、18、20、23、26、29号CpG位点在≥50岁年龄组的甲基化水平均高于<50岁年龄组（均P<0.05）。肝细胞癌中ChREBP基因mRNA表达水平低于癌旁组织（P = 0.003），但与甲基化无显著相关性（P<0.05）。
结论肝细胞癌中ChREBP基因mRNA表达水平低于癌旁组织，但这一改变可能不是通过影响DNA的甲基化水平实现的。

Abstract:
Objective To detect the expression of ChREBP mRNA and its CpG island methylation in human hepatocellular carcinoma tissues and investigate their correlation.
Methods Bisulfite Genomic sequencing was used to analyze the methylation level of 29 CpG sites in CpG island of ChREBP in 45 paired hepatocellular carcinoma and adjacent non-tumor tissues. The expression of ChREBP mRNA was detected in 31 paired hepatocellular carcinoma and adjacent non-tumor tissues using quantitative real-time PCR, meanwhile, the relationship between the level of methylation status and mRNA expression was analyzed.
Results The methylation level of some CpG sites (5, 6, 7, 14) were significantly lower in hepatocellular carcinoma tissues than those in the adjacent non-tumor tissues (P<0.05). There was no statistical significancet difference in the methylation level of other CpG sites between hepatocellular carcinoma tissues and adjacent non-tumor tissues (all P<0.05). The methylation level of ChREBP at CpG 15, 18, 20, 23, 26 and 29 tended to be higher in patients with older age (≥50 years) than those in younger age(<50 years) (all P<0.05). The mRNA expression of ChREBP in hepatocellular carcinoma tissues was lower than that in adjacent non-tumor tissues (P=0.003), however, no significant relationship was observed between DNA methylation and the mRNA expression of ChREBP in hepatocellular carcinoma tissues(P<0.05).
Conclusion There was a down-regulated expression of ChREBP in hepatocellular carcinoma tissues, which could not be mediated by DNA methylation.

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