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目的评估外周血浆DNA甲基化检测在肝细胞肝癌(HCC)诊断中的作用,筛选血浆灵敏度及特异性高的甲基化抑癌基因。方法采用甲基化特异性PCR(MSP)检测34例经病理检查证实的HCC患者血浆及其配对癌组织中的SLIT2及DAPK基因启动子甲基化状态,并分析其与HCC临床病理特征的关系。结果 34例HCC患者癌组织中SLIT2和DAPK基因甲基化率分别为70.6%(24/34)和79.4%(27/34),相应外周血浆中SLIT2和DAPK基因甲基化率分别为44.1%(15/34)和50.0%(17/34)。血浆SLIT2和DAPK基因甲基化检测的灵敏度分别为62.5%和63.0%,特异度均为100%,阴性预测值分别为52.6%和41.2%,阳性预测值均为100%。HCC癌组织和外周血浆中SLIT2及DAPK甲基化检出率与HCC各临床病理特征无关(P>0.05)。在AFP<400μg/L患者中SLIT2和DAPK基因甲基化联合检出率达61.1%(11/18)。结论基于MSP方法,血浆中SLIT2及DAPK基因甲基化仍有较高的检出率,HCC癌组织和血浆中SLIT2及DAPK基因甲基化具有明显的一致性。血浆DNA甲基化可作为一种独立非侵入性诊断HCC的标志物,尤其对AFP阴性者可提高HCC的诊断率。HBV感染可能仅与部分抑癌基因甲基化相关。
Objective To evaluate the role of peripheral plasma DNA methylation in the diagnosis of hepatocellular carcinoma (HCC) and to screen the methylation suppressor gene with high sensitivity and specificity. Methods Methylation-specific PCR (MSP) was used to detect the methylation status of SLIT2 and DAPK promoter in plasma and matched cancer tissues of 34 HCC patients confirmed by pathology. The relationship between the methylation status and the clinicopathological features of HCC . Results The methylation rates of SLIT2 and DAPK gene in 34 HCC patients were 70.6% (24/34) and 79.4% (27/34), respectively. The methylation rates of SLIT2 and DAPK genes in the corresponding peripheral blood plasma were 44.1% (15/34) and 50.0% (17/34) respectively. The sensitivity of plasma methylation of SLIT2 and DAPK genes were 62.5% and 63.0% respectively, specificity was 100%, negative predictive value was 52.6% and 41.2% respectively, and the positive predictive value was 100%. The detection rate of methylation of SLIT2 and DAPK in HCC tissues and peripheral plasma was not related to the clinicopathological features of HCC (P> 0.05). The methylation detection rate of SLIT2 and DAPK gene in AFP <400μg / L patients was 61.1% (11/18). Conclusion Based on the MSP method, the methylation of SLIT2 and DAPK genes in plasma is still high, and the methylation of SLIT2 and DAPK genes in HCC tissues and plasma has obvious consistency. Plasma DNA methylation can be used as a marker for independent noninvasive diagnosis of HCC, especially for patients with AFP that can improve the diagnosis of HCC. HBV infection may be related only to partial tumor suppressor gene methylation.