PKM2在肝内胆管癌中的表达及其致癌作用

发布时间：2018-06-07 08:08

本文选题：肝内胆管癌 + PKM2　；参考：《浙江大学》2015年硕士论文

【摘要】：背景肝内胆管癌(ICC)是消化系统中的一种相对少见而且最具破坏性的恶性肿瘤之一,具有发生隐匿、恶性程度高、发展迅速、预后较差等特点,而关于肝内胆管癌的研究却相对较少,其发生发展机制尚不明确；丙酮酸激酶M2(PKM2)已被报道参与有氧和无氧糖酵解,并且已被证实和多种肿瘤发生和进展相关。然而,在肝内胆管癌组织中尚无关于PKM2的报道。本研究的目的是探讨PKM2在肝内胆管癌中的表达以及它肝内胆管癌发生发展中的作用。方法收集50例肝内胆管癌组织及其配对的癌旁组织,采用RT-PCR和免疫组织化学的方法检测PKM2在肝内胆管癌癌组织和配对癌旁组织中的表达水平,并分析其表达水平与肝内胆管癌远期生存预后的关系；构建PKM2的干扰RNA并转染9810和CCLP-1细胞系,检测PKM2蛋白的表达水平变化,并观察对比9810和CCLP-1细胞侵袭、迁移和增殖能力的变化。结果 RT-PCR结果提示肝内胆管癌组织mRNA相对表达水平为3.24±0.693,显著高于相对应的癌旁组织的表达水平0.48±0.109,差异有统计学意义(P0.001)；免疫组化的结果表明:PKM2低表达组(评分≤7分)平均生存期为40.61±4.537月,明显高于PKM2高表达组(评分7分)的15.15±2.368月,差异有统计学意义(P0.001)；在9810和CCLP-1细胞系中的,干扰组与对照组相比,在侵袭和转移实验中,对照组细胞侵袭及迁移计数显著高于干扰组(P0.001)；在增殖实验中随着时间增长,在第48h、72h对照组的吸光度值显著高于干扰组(P0.001)。结论 PKM2在肝内胆管癌组织中高表达,并在肝内胆管癌细胞的侵袭、迁移和增殖过程中起着重要作用,其表达程度高低还和肝内胆管癌患者术后长期预后密切相关；PKM2可作为肝内胆管癌诊断、靶向治疗及判断预后的潜在重要靶标。
[Abstract]:Background Intrahepatic cholangiocarcinoma (ICC) is one of the relatively rare and most destructive malignant tumors in the digestive system, with the characteristics of occult occurrence, high degree of malignancy, rapid development and poor prognosis. The pathogenesis of pyruvate kinase M2-PKM2 has been reported to be involved in aerobic and anaerobic glycolysis and has been proved to be related to the occurrence and progression of various tumors. However, PKM2 has not been reported in intrahepatic cholangiocarcinoma. The purpose of this study was to investigate the expression of PKM2 in intrahepatic cholangiocarcinoma and its role in the development of intrahepatic cholangiocarcinoma. Method Fifty cases of intrahepatic cholangiocarcinoma tissues and their matched adjacent tissues were collected. The expression of PKM2 in intrahepatic cholangiocarcinoma tissues and matched paracancerous tissues was detected by RT-PCR and immunohistochemistry. The relationship between the expression of PKM2 protein and the long-term survival and prognosis of intrahepatic cholangiocarcinoma was analyzed, the interference RNA of PKM2 was constructed and transfected into 9810 and CCLP-1 cell lines, the expression of PKM2 protein was detected, and the invasion of 9810 and CCLP-1 cells was observed and compared. Changes in migration and proliferation. Result The results of RT-PCR showed that the relative expression level of mRNA in intrahepatic cholangiocarcinoma was 3.24 卤0.693, which was significantly higher than that in the corresponding paracancerous tissues (0.48 卤0.109). The mean survival time was 40.61 卤4.537 months. It was significantly higher than that in the high expression group of PKM2 (score 7) (15.15 卤2.36 months), and the difference was statistically significant (P 0.001). In the 9810 and CCLP-1 cell lines, the invasion and metastasis of the interference group were compared with those of the control group. The number of cell invasion and migration in the control group was significantly higher than that in the interference group (P 0.001), and the absorbance value in the control group was significantly higher than that in the interference group at 48 h and 72 h with the increase of time. Conclusion PKM2 is highly expressed in intrahepatic cholangiocarcinoma and plays an important role in invasion, migration and proliferation of intrahepatic cholangiocarcinoma cells. The level of PKM2 expression is closely related to the long-term prognosis of patients with intrahepatic cholangiocarcinoma. PKM2 can be used as a potential target for the diagnosis, targeted therapy and prognosis of intrahepatic cholangiocarcinoma.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.8

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