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背景与目的:DNA甲基化是一种新的肿瘤诊断及预后判断的标志物。本研究运用自行建立的甲基化敏感性限制性内切酶-定量PCR(methylation-sensitive restriction enzymes-based quantitative PCR,MSRE-qPCR)方法检测血浆GSTP1和SFRP1基因的DNA甲基化状态,探讨其在肝细胞癌(hepatocellular carcinoma,HCC)早期诊断中的价值。方法:收集150例血浆标本,包括72例HCC,37例肝良性病变和41名健康对照者。用MSRE-qPCR法检测血浆GSTP1和SFRP1基因DNA甲基化水平。结果:HCC患者血浆GSTP1和SFRP1甲基化阳性率分别为54.2%和27.8%,显著高于健康对照组(9.8%和2.4%,P<0.001)和肝良性病变组(10.8%和5.4%,P<0.001);同时检测血浆GSTP1和SFRP1可检出63.9%的HCC;而联合血清AFP分析可进一步将HCC诊断率提高至73.6%。结论:联合检测血浆GSTP1和SFRP1基因DNA甲基化对于HCC早期非侵入性诊断具有重要价值。
BACKGROUND & OBJECTIVE: DNA methylation is a new marker of tumor diagnosis and prognosis. In this study, methylation-sensitive restriction enzymes-based quantitative PCR (MSRE-qPCR) was used to detect DNA methylation status of plasma GSTP1 and SFRP1 genes, In the early diagnosis of hepatocellular carcinoma (HCC). Methods: 150 plasma samples were collected, including 72 HCCs, 37 benign liver lesions and 41 healthy controls. The levels of DNA methylation in plasma GSTP1 and SFRP1 were detected by MSRE-qPCR. Results: The positive rates of GSTP1 and SFRP1 methylation in HCC patients were 54.2% and 27.8% respectively, which were significantly higher than those in healthy controls (9.8% vs 2.4%, P <0.001) and benign liver lesions (10.8% vs 5.4% P <0.001). Simultaneous detection of plasma GSTP1 and SFRP1 detected 63.9% of HCC. Combined serum AFP analysis further increased the diagnostic yield of HCC to 73.6%. Conclusion: Combined detection of plasma DNA methylation of GSTP1 and SFRP1 genes is of great value in the early noninvasive diagnosis of HCC.