肝细胞癌中FHIT基因启动子甲基化状态分析及其与临床病理特征之间的关系研究

Analysis of the Status of FHIT Gene Promoter Methylation and Its Association with Clinicopathological Features of Patients with Hepatocelluar Carcinoma

目的:分析肝细胞癌组织中FHIT基因启动子甲基化状态及其与FHIT基因表达和肝细胞癌临床病理特征的关系。方法:运用甲基化特异性PCR(MSP)方法分析肝细胞癌组织和癌旁正常肝脏组织中FHIT基因启动子甲基化状况;应用RT-PCT和Western免疫印迹方法检测FHIT基因mRNA和蛋白的表达情况;统计学分析FHIT基因启动子甲基化与肝细胞癌临床病理特征的关系。结果:MSP分析结果表明肝细胞癌组织中FHIT基因甲基化率(60.8%)显著高于癌旁正常组织中FHIT基因甲基化(16.2%;x2=31.071,P=0.000)。同时我们还发现:发生完全或者部分甲基化的肝细胞癌组织或者癌旁正常肝组织中FHIT基因mRNA和蛋白表达水平显著降低。FHIT基因启动子甲基化和肝细胞癌患者的临床分期和肝外转移情况密切相关(P=0.006和0.049),而与其他临床病理特征无相关性(P>0.05)。结论:FHIT基因甲基化是导致FHIT基因在肝细胞癌中失活的一个重要因素,与肝细胞癌的发生密切相关,有望成为肝细胞癌早期诊断的分子检测标志物和分子治疗新靶点。

Objective：To analyse of the status of FHIT promoter methylation and its association with FHIT gene expression and clinicopathological features of patients with hepatocelluar carcinoma（HCC） . Methods：Methylation-specific PCR assay was performed to detect the status of FHIT promoter methylation in HCC tissues and corresponding non-tumor tissues. RT-PCR and Western blotting assays were performed to detect the expression of FHIT mRNA and protein. Statistical analyses were performed to analyze the association between FHIT promoter methylation and clinicopathological features of HCC patients. Results：The results of MSP assays showed that the rate of FHIT promoter methylation in HCC tissues（60.8%） was significantly higher than that in corresponding non-tumor tissues（16.2%;x2=31.071,P=0.000） . Moreover,the expression of FHIT mRNA and protein in HCC or corresponding non-tumor tissues with full or partial methylation was significantly lower than that in those tissues with unmethylation. Additionally,statistical analyses showed that the FHIT promoter methylation was significantly associated with clinical stage and metastasis of HCC patients（P=0.006 and 0.049,respectively,but not with other features of HCC patients（P0.05） . Conclusion：The promoter methylation of FHIT gene might be an important factor for inactivation of FHIT gene in HCC. FHIT gene methylation was associated with hepatocarcinoma and would be molecular marker and target for early detection and therapy of HCC.