肝癌患者血浆RASSF1A异常甲基化检测的意义

目的:采用甲基化特异性PCR法(MSP)检测41例原发性肝癌(HCC)患者血浆中游离Ras相关结构域家族蛋白1A(RASSF1A)异常高甲基化情况,探讨血浆RASSF1A检测在肝癌诊断中的意义. 方法:收集患者血浆,提取游离DNA,亚硫酸氢钠修饰并纯化,以甲基化特异性引物及非甲基化引物进行PCR. 结果: 41例患者血浆中有17例RASSF1A呈现异常高甲基化(41.5%),对照血浆均为阴性. 患者一般情况、临床病理资料等与血浆中RASSF1A异常高甲基化检测结果无关(P＞0.05);在10例AFP阴性的HCC患者中,有7例结果为阳性. 结论:血浆RASSF1A甲基化检测对肝癌的早期诊断及筛选有重要意义,并有助于判断预后.

Diagnostic value of detection of aberrantly methylated RASSF1A in plasma of patients with hepatocellular carcinoma

AIM: To study the aberrant methylation status of Ras association domain family 1A (RASSF1A) in the plasma of patients with hepatocellular carcinoma (HCC) by methylation-specific PCR (MSP),and to explore the value of circular tumor-related DNA in diagnosis of HCC. METHODS: Free DNA was extracted from peripheral blood plasma of 41 HCC patients. After modified with sodium bisulfite and purified,the DNA was amplified using primers specific for methylated and unmethylated sequences of RASSF1A. RESULTS: Aberrant RASSF1A methy- lation was found in 41.5% (17 of 41) of plasma DNA in HCC patients. No methylated RASSF1A was detected in 10 cases of control plasma samples. Clinical data including age,sex,clinical stages,levels of AFP,hepatitis status,number of tumor,invasion,metastasis and cirrhosis showed no correlation with aberrant methylation of RASSF1A (P>0.05). Aberrantly methylated RASSF1A was also detected in 7 of 10 plasma samples of AFP negative cases. CONCLUSION: Detection of aberrant methylation of RASSF1A in blood is feasible to early detect and screen HCC,and is useful to estimate the prognosis.