CAJ | 학술논문

目的：建立实时荧光定量甲基化方法研究肝细胞癌（HCC）患者外周血血清中游离谷胱甘肽-硫-转移酶P1基因（GSTP1）启动子区域甲基化状态，探讨其是否可作为HCC早期诊断的指标。方法收集95例血清标本，包括40例HCC、30例肝硬化和25例健康体检者，采用实时荧光定量甲基化特异性聚合酶链反应（PCR）技术对血清中GSTP1基因的甲基化水平进行定量分析，应用接受者操作特性（ROC）曲线评估其诊断价值。结果肝癌患者血清中GSTP1基因甲基化定量水平明显高于健康对照组，差异有统计学意义（P＜0．05）。ROC曲线分析表明通过GSTP1基因甲基化定量分析能够高效区分肝癌和肝硬化及健康者（AUC＝0．8641），以甲基化率2％作为诊断HCC的临界值，其诊断特异度为87．5％，敏感度达69．6％；联合检测血清GSTP1基因甲基化和血清AFP可将HCC检出率提高至75％。结论实时荧光定量甲基化方法可精确定量血清中GSTP1基因甲基化水平，在HCC的早期诊断中有一定的应用价值。
목적：건립실시형광정량갑기화방법연구간세포암（HCC）환자외주혈혈청중유리곡광감태-류-전이매P1기인（GSTP1）계동자구역갑기화상태，탐토기시부가작위HCC조기진단적지표。방법수집95례혈청표본，포괄40례HCC、30례간경화화25례건강체검자，채용실시형광정량갑기화특이성취합매련반응（PCR）기술대혈청중GSTP1기인적갑기화수평진행정량분석，응용접수자조작특성（ROC）곡선평고기진단개치。결과간암환자혈청중GSTP1기인갑기화정량수평명현고우건강대조조，차이유통계학의의（P＜0．05）。ROC곡선분석표명통과GSTP1기인갑기화정량분석능구고효구분간암화간경화급건강자（AUC＝0．8641），이갑기화솔2％작위진단HCC적림계치，기진단특이도위87．5％，민감도체69．6％；연합검측혈청GSTP1기인갑기화화혈청AFP가장HCC검출솔제고지75％。결론실시형광정량갑기화방법가정학정량혈청중GSTP1기인갑기화수평，재HCC적조기진단중유일정적응용개치。
Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .