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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 expression and significance of pyruvate kinase M2 (PKM2) and yes-related protein (YAP) in hepatocellular carcinoma (HCC) and to explore the correlation between PKM2 protein and YAP protein expression in hepatocellular carcinoma. Methods Retrospectively collected 120 cases of hepatocellular carcinoma and their peritumoral tissue resected from April 2010 to October 2013 in the Fourth Affiliated Hospital of China Medical University. SP immunohistochemistry was used to detect 120 cases of hepatocellular carcinoma and carcinoma Adjacent tissues PKM2 and YAP protein expression and analysis of the relationship between the expression of PKM2 protein and YAP protein in hepatocellular carcinoma and clinicopathological features. Among them, 50 cases of HCC and paracancerous tissue samples were taken fresh frozen tissue specimens immediately surgery, set the liquid nitrogen cooled and sent to -70 ° C freezer. Western blot and real-time fluorescence quantitative PCR were used to detect the relative expression levels of PKM2, YAP protein and RNA in 50 cases of HCC and paracancerous tissues respectively. Results1 Immunohistochemistry results showed that the positive expression rate of PKM2 protein was 67.50% (81/120) and the positive expression rate of YAP protein was 71.67% (86/120) in hepatocellular carcinoma tissues, which were higher than that in the adjacent tissues [PKM2 Protein: 20.83% (25/120); YAP protein: 29.17% (35/120)], the difference was statistically significant (P <0.050). There was a positive correlation between PKM2 protein and YAP protein expression in HCC (r = 0.519, P <0.001). The expression of PKM2 protein in HCC tissues correlated with tumor diameter, TNM stage, tumor differentiation and AFP level (P <0.050). The expression of YAP protein was correlated with tumor differentiation and AFP level (P < 0.050). The results of Western blot showed that the expression levels of PKM2 and YAP in hepatocellular carcinoma tissues were 1.25 ± 0.11 and 1.08 ± 0.10, respectively, and those in paracancerous tissues were 0.38 ± 0.01 and 0.41 ± 0.02, respectively. The expressions of PKM2 and YAP in hepatocellular carcinoma The levels were higher than the adjacent tissues, the difference was statistically significant (P <0.050). The results of real-time fluorescence quantitative PCR showed that the expression levels of PKM2 m RNA and YAP m RNA in HCC tissues were 11.38 ± 0.35 and 19.96 ± 0.48, respectively, which were higher than those in paracancerous tissues, The differences were statistically significant (P <0.050). Conclusion PKM2 protein and YAP protein are correlated with the occurrence of HCC and are closely related to the differentiation of HCC.