PKM2和YAP在人肝癌组织中的表达及其相关性分析

The Expression and Correlation of PKM2 and YAP in Hepatocellular Carcinoma Tissue

目的探讨丙酮酸激酶M2(PKM2)和yes相关蛋白(YAP)在肝癌组织中的表达及其意义,并探索肝癌组织中PKM2蛋白和YAP蛋白表达的相关性。方法回顾性收集2010年4月至2013年10月期间于中国医科大学附属第四医院行手术切除的120例肝癌及其癌旁组织标本,采用SP免疫组织化学染色方法检测120例肝癌组织及癌旁组织PKM2和YAP蛋白的表达,并分析肝癌组织中PKM2蛋白和YAP蛋白表达与临床病理学特征的关系。其中,50例肝癌及癌旁组织标本具备手术时立即取材的新鲜冰冻组织标本,置液氮冷却后送-70°C冷冻冰箱内保存。采用Western blot与实时荧光定量PCR方法分别检测50例肝癌组织及癌旁组织中PKM2、YAP蛋白及其RNA的相对表达水平。结果 1免疫组化染色结果显示:肝癌组织中PKM2蛋白的表达阳性率为67.50%(81/120),YAP蛋白的表达阳性率为71.67%(86/120),均高于癌旁组织〔PKM2蛋白:20.83%(25/120);YAP蛋白:29.17%(35/120)〕,差异均有统计学意义(P<0.050)。2肝癌组织中,PKM2蛋白和YAP蛋白的表达呈正相关(r=0.519,P<0.001)。3肝癌组织中PKM2蛋白的表达与肿瘤直径、TNM分期、肿瘤分化程度及甲胎蛋白(AFP)水平均相关(P<0.050),YAP蛋白的表达与肿瘤分化程度和AFP水平均有关(P<0.050)。4 Western blot结果显示,肝癌组织中PKM2蛋白和YAP蛋白的表达水平分别为1.25±0.11和1.08±0.10,癌旁组织分别为0.38±0.01和0.41±0.02,肝癌组织中PKM2蛋白和YAP蛋白的表达水平均高于癌旁组织,差异均有统计学意义(P<0.050)。5实时荧光定量PCR结果表明,以癌旁组织作为基准(即为1),肝癌组织中PKM2 m RNA和YAP m RNA的表达水平分别为11.38±0.35和19.96±0.48,均高于癌旁组织,差异均有统计学意义(P<0.050)。结论 PKM2蛋白和YAP蛋白与肝癌的发生相关,并均与肝癌的分化程度密切相关。

Objective To investigate the significance of expression and correlation of pyruvate kinase M2 （PKM2） and yes association protein （YAP） in hepatocellular carcinoma （HCC） tissue, and then explore the relationship between the 2 kinds of protein. Methods A total of 120 patients＇ HCC tissues and adjacent tissues were collected retrospectively, who treated in our hospital from Apr. 2010 to Oct. 2013, the expressions of PKM2 and YAP protein in these HCC tissues and adjacent tissues were detected by SP immunohistochemical method, and then analyzed the relationship between the expressions of PKM2 and YAP ptotein with the clinicopathological features of HCC. Of the 120 patients, the expressions of YAP and PKM2 protein and its mRNA in 50 cases of HCC tissues and adjacent tissues were also examined by Western blot and real-time PCR methods respectively. Results （1） The immunohistochemical results showed that, the positive rate of PKM2 protein and YAP protein in HCC tissues were 67.50% （81/120） and 71.67% （86/120） respectively, which were both higher than those of adjacent tissues [PKM2 protein： 20.83% （25/120）; YAP protein： 29.17% （35/120）],P〈0.050. （2） The expression of PKM2 protein was significantly positively correlated with the expression of YAP protein in HCC tissues （r=0.519, P〈0.001）. （3） In HCC tissues, the expression of PKM2 protein was significantly correlated with the diameter of tumor, TNM staging, differentiation of HCC, and lever of alpha fetal protein （P〈0.050）, and the expression of YAP protein was significantly correlated with the differentiation of HCC and lever of alpha fetal protein （P〈0.050）. （4） Western blot results showed that, the expression levels of PKM2 protein and YAP protein in HCC tissues were 1.25± 0.11 and 1.08±0.10 respectively, which were significantly higher than those of ad）acent tissues （PKM2 protein： 0.38±0.01, YAP protein： 0.41±0.02）, P〈0.050. （5） Real-time PCR assays results showed that, basing the expressions of PKM2 mRNA and YAP mRNA in adjacent tissues （both as 1）, expressions of PKM2 mRNA and YAP mRNA in HCC tissues were 11.38±0.35 and 19.96±0.48 respectively, which were both higher than those of adjacent tissues （P〈0.050）. Conclusion PKM2 and YAP protein were related to the initiation of HCC, and they were also closely correlated with the differentiation and prognosis of HCC.