肝癌患者血浆RASSF1A与APC基因甲基化诊断和预后价值分析

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目的:探讨肝癌患者血浆中抑癌基因RASSF1A与APC启动子区域甲基化与诊断及生存期预后的关系。方法:收集102例肝癌、80例肝硬化及100名健康人血浆,提取DNA,采用甲基化特异性聚合酶链反应法检测RASSF1A与APC基因启动子区域甲基化状态,随访其生存期并分析基因甲基化状态与肝癌诊断及预后的关系。结果:RASSF1A基因启动子区域甲基化阳性率在肝癌和肝硬化血浆中分别为51.96%(53/102)和15.00%(12/80),差异有统计学意义,χ2=26.677,P=0.000;APC基因启动子区域甲基化阳性率在肝癌和肝硬化血浆中分别为47.06%(48/102)和22.50%(18/80),差异有统计学意义,χ2=11.700,P=0.001。102例肝癌患者中,44例(43.14%)AFP检测为阳性;58例AFP阴性的患者中,RASSF1A基因和APC基因启动子区域甲基化的阳性率分别为53.45%(31/58)和46.55%(27/58)。RASSF1A基因启动子区域甲基化阳性组与阴性组生存期比较,差异有统计学意义,P=0.011;而APC基因阳性组与阴性组生存期比较,差异无统计学意义,P=0.175。Cox多因素分析结果显示,AFP、临床分期、治疗方式及门静脉瘤栓有无为肝癌独立预后因子,RASSF1A和APC基因同时甲基化是肝癌的独立预后因子,P=0.019。结论:RASSF1A与APC基因启动子区域甲基化对肝癌、特别是AFP阴性肝癌的诊断有帮助,RASSF1A和APC基因启动子区域同时甲基化可以作为评价肝癌生存期预后的独立预后因子。 Objective: To investigate the relationship between the methylation status of RASSF1A and APC promoter and the prognosis of patients with liver cancer. Methods: DNA was extracted from 102 cases of hepatocellular carcinoma, 80 cases of liver cirrhosis and 100 healthy human plasma. The methylation status of RASSF1A and APC gene promoter region was detected by methylation specific polymerase chain reaction Analysis of the relationship between gene methylation status and diagnosis and prognosis of liver cancer. Results: The methylation positive rate of RASSF1A gene promoter region was 51.96% (53/102) and 15.00% (12/80) in liver cancer and cirrhosis plasma respectively, the difference was statistically significant (χ2 = 26.677, P = 0.000 The positive rate of APC gene promoter methylation in hepatocellular carcinoma and cirrhosis was 47.06% (48/102) and 22.50% (18/80), respectively. The difference was statistically significant (χ2 = 11.700, P = 0.001). The positive rate of AFP was 44 (43.14%) in 102 cases of hepatocellular carcinoma. The positive rate of methylation of RASSF1A gene and APC gene in 58 AFP negative patients was 53.45% (31/58) and 46.55 % (27/58). The difference was statistically significant (P = 0.011) between the methylation-positive group and the negative group in the RASSF1A gene promoter region, but there was no significant difference in the survival time between the APC gene positive group and the negative group (P = 0.175). Cox multivariate analysis showed that AFP, clinical stage, treatment and portal vein tumor thrombus were independent prognostic factors for HCC. Simultaneous methylation of RASSF1A and APC genes was an independent prognostic factor for HCC (P = 0.019). CONCLUSION: Methylation of RASSF1A and APC gene promoters is helpful for the diagnosis of liver cancer, especially AFP negative liver cancer. Simultaneous methylation of RASSF1A and APC promoter regions may be an independent prognostic factor for prognosis of liver cancer.
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