肝癌中CpG甲基化与microRNA-34沉默的研究

Study on Methylation of MicroRNA-34 in the Hepatocellular Carcinoma

目的探讨肝癌中miR-34家族甲基化的价值。方法入组经病理组织学确诊的43例原发性肝癌手术病例以及临近癌旁组织,使用甲基化特异性PCR(MSP)方法分析miR-34在肝癌组织和癌旁组织中的甲基化状态,使用逆转录PCR法验证MSP结果。结果肝癌组织中miR-34a和miR-34b/c甲基化频率启动分别为72.1%和79.1%,均高于癌旁组织(P<0.05);miR-34a和miR-34b/c在肝癌组织中明显比癌旁组织表达下调(P<0.05);肝癌组织中miR-34b表达与CpG甲基化呈负相关(P<0.05)。结论肝癌组织中DNA甲基化可能参与了miR-34b的失活。

Objective To explore the value of methylation of microRNA-34 （miR-34） family in the hepatocellular carcinoma. Methods Tumor and adjacent non-tumor tissues were collected from 43 hepatoeellular carcinoma patients who underwent surgery. Methylation specific PCR （MSP） method was used to analyze the methylation status of miR-34 in the hepatoeellular carcinoma tissues and the adjacent non-tumor tissues, and the results were verified by reverse transcription PCR method. Results The methylation frequencies of miR-34a and miR-34b/c were 72.1% and 79.1% in the hepatoeellular carcinoma tissues,which were significantly higher than that in the adjacent non-tumor tissues （ P 〈 0.05 ） , respectively. The expressions of miR-34a and miR-34b/c were significantly downregulated in the hepatocellular carcinoma tissues compared with adjacent non-tumor tissues （P 〈0.05 ）. The expression of miR-34b were negatively related with DNA methylation in the hepatocellular carcinoma tissues. Conclusion DNA methylation may be involved in the inactivation of miR-34b in HCC.