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目的探讨高尔基体蛋白73(GP73)与RASSF1A基因启动子区甲基化对原发性肝癌(HCC)的诊断价值。方法选择2012年8月-2014年2月在浙江省宁波市第六医院住院的80例肝癌患者、40例肝硬化、30例健康体检者,采用酶联免疫法(ELISA)检测血清中GP73含量,甲基化特异性PCR(methylation specific PCR,MSP)检测血清中RASSF1A基因启动子区甲基化情况,ROC曲线分析GP73对HCC的最佳诊断点,并对单一及组合(GP73+RASSF1A)方式的敏感度、特异度进行比较。结果肝癌组、肝硬化组、健康体检组血清GP73平均浓度为278.3μg/L、187.3μg/L、46.8μg/L(F=6.85,P<0.05);RASSF1A基因甲基化检出率分别为65%(52/80)、20%(8/40)、3.3%(1/30)(χ2=44.1,P<0.05);GP73诊断肝癌的ROC曲线下面积为0.852,P<0.05,取GP73为122.02μg/L为最佳诊断分界点;GP73诊断HCC的灵敏度高于RASSF1A基因甲基化,GP73诊断HCC的特异度低于RASSF1A基因甲基化,GP73与RASSF1A基因甲基化并联诊断HCC的敏感度与单项GP73相比,差异具有统计学意义(χ2=4.12,P<0.05),两项指标并联诊断HCC的特异度与单项RASSF1A甲基化比较,差异无统计学意义(χ2=0.31,P>0.05)。结论 GP73并联RASSF1A基因甲基化检测对诊断HCC的敏感度较高,具有一定的筛查能力。
Objective To investigate the value of methylation of Golgi apparatus protein 73 (GP73) and RASSF1A promoter in the diagnosis of primary hepatocellular carcinoma (HCC). Methods Eighty patients with HCC, 40 patients with cirrhosis and 30 healthy individuals admitted to the Sixth Hospital of Ningbo City, Zhejiang Province from August 2012 to February 2014 were enrolled in this study. Serum GP73 levels were determined by enzyme-linked immunosorbent assay (ELISA) Methylation specific PCR (MSP) was used to detect the promoter methylation status of RASSF1A gene in serum. ROC curve was used to analyze the best diagnostic point of GP73 for HCC. The single and combined (GP73 + RASSF1A) The sensitivity and specificity were compared. Results The average serum concentrations of GP73 in HCC group, cirrhosis group and healthy group were 278.3μg / L, 187.3μg / L and 46.8μg / L respectively (F = 6.85, P <0.05). The methylation rates of RASSF1A gene were The area under the ROC curve of GP73 in the diagnosis of HCC was 0.852 (P <0.05), and GP73 The sensitivity of GP73 for diagnosis of HCC was higher than that of RASSF1A gene, the specificity of GP73 for diagnosis of HCC was lower than that of RASSF1A gene, and the methylation of GP73 and RASSF1A gene for diagnosis of HCC The sensitivity was significantly different from that of single GP73 (χ2 = 4.12, P <0.05). There was no significant difference in the specificity between the two indexes in diagnosing HCC and the single RASSF1A methylation (χ2 = 0.31, P> 0.05). Conclusion The methylation detection of GP73 in parallel with RASSF1A gene is more sensitive to diagnose HCC and has certain screening ability.