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目的研究慢性乙型病毒性肝炎(乙肝)肝硬化患者外周血甲胎蛋白(AFP)、丙酮酸激酶M2(PKM2)、白蛋白(Alb)mRNA表达及其临床意义。方法选择89例慢性乙肝肝硬化患者作为肝硬化组,另选择60例健康人群作为对照组。肝硬化组按照血清HBV-DNA检测结果,再分为病毒复制组53例和非病毒复制组36例。抽取各组外周血5 mL,实时定量PCR法检测肝硬化组与对照组血浆AFP、PKM2、Alb mRNA表达水平。荧光定量PCR检测肝硬化患者血清HBV-DNA含量,分析血浆AFP、PKM2、Alb mRNA与HBV-DNA载量的相关性。根据AFP、PKM2、Alb mRNA相对表达量,绘制相应ROC曲线。结果肝硬化组AFP mRNA相对表达量为13.6±2.1,PKM2 mRNA为6.4±0.5,Alb mRNA为15.1±2.5。对照组分别为7.6±0.7,1.4±0.2,7.2±0.5,肝硬化组AFP、PKM2、Alb mRNA表达量显著高于对照组,差异有统计学意义(P<0.05)。病毒复制组AFP mRNA相对表达量为15.3±4.2,PKM2 mRNA为7.6±0.7,Alb mRNA为18.2±4.5。非病毒复制组分别为11.1±2.4,4.6±0.3,10.5±1.7,病毒复制组AFP、PKM2、Alb mRNA表达量显著高于非病毒复制组,差异有统计学意义(P<0.05)。相关性分析显示,AFP mRNA与HBV-DNA呈正相关(r=0.401,P<0.05),PKM2 mRNA与HBV-DNA呈正相关(r=0.380,P<0.05),Alb mRNA与HBV-DNA呈正相关(r=0.607,P<0.001)。AFP mRNA ROC曲线下面积为0.885,最佳临界值为8.7,对应的敏感度为84.9%,特异度为84.3%;PKM2 mRNA ROC曲线下面积为0.879,最佳临界值为3.4,对应的敏感度为84.1%,特异度为81.7%;Alb mRNA ROC曲线下面积为0.921,最佳临界值为9.5,对应的敏感度为88.1%,特异度为96.8%。结论肝硬化患者血浆AFP、PKM2、Alb mRNA表达显著增加,通过检测外周血AFP、PKM2及Alb mRNA可以为肝硬化诊断提供参考。
Objective To study the expression of AFP, PKM2 and Alb in patients with chronic hepatitis B (HBV) cirrhosis and its clinical significance. Methods Eighty-nine patients with chronic hepatitis B cirrhosis were selected as the cirrhosis group and another 60 healthy people as the control group. According to the results of serum HBV-DNA test, cirrhosis group was further divided into 53 cases of virus replication group and 36 cases of non-virus replication group. 5 mL of peripheral blood was drawn from each group, and the levels of AFP, PKM2 and Alb mRNA in cirrhosis and control groups were detected by real-time quantitative PCR. Fluorescent quantitative PCR was used to detect serum HBV-DNA in patients with cirrhosis. The correlation between plasma AFP, PKM2, Alb mRNA and HBV-DNA load was analyzed. According to the relative expression of AFP, PKM2, Alb mRNA, draw the corresponding ROC curve. Results The relative expression of AFP mRNA in liver cirrhosis group was 13.6 ± 2.1, PKM2 mRNA was 6.4 ± 0.5, Alb mRNA was 15.1 ± 2.5. The control group were 7.6 ± 0.7, 1.4 ± 0.2 and 7.2 ± 0.5 respectively. The expression of AFP, PKM2 and Alb in cirrhosis group was significantly higher than that in control group (P <0.05). The relative expression of AFP mRNA in viral replication group was 15.3 ± 4.2, PKM2 mRNA was 7.6 ± 0.7, Alb mRNA was 18.2 ± 4.5. The levels of AFP, PKM2 and Alb mRNA in viral replication group were significantly higher than those in non-virus replication group (P <0.05). Correlation analysis showed that there was a positive correlation between AFP mRNA and HBV-DNA (r = 0.401, P <0.05), PKM2 mRNA and HBV DNA (r = 0.380, r = 0.607, P <0.001). The area under the ROC curve of AFP mRNA was 0.885, the best cut-off value was 8.7, the corresponding sensitivity was 84.9% and the specificity was 84.3%. The area under the ROC curve of PKM2 mRNA was 0.879 and the best cut-off value was 3.4. The corresponding sensitivity And the specificity was 81.7%. The area under the Alb mRNA ROC curve was 0.921, the best cut-off value was 9.5, corresponding to a sensitivity of 88.1% and a specificity of 96.8%. Conclusions The expression of AFP, PKM2 and Alb mRNA in cirrhotic patients is significantly increased. The detection of AFP, PKM2 and Alb mRNA in peripheral blood can provide reference for the diagnosis of cirrhosis.