本文選題：肝內(nèi)膽管癌 + PKM2��； 參考：《浙江大學(xué)》2015年碩士論文

【摘要】：背景 肝內(nèi)膽管癌(ICC)是消化系統(tǒng)中的一種相對(duì)少見而且最具破壞性的惡性腫瘤之一,具有發(fā)生隱匿、惡性程度高、發(fā)展迅速、預(yù)后較差等特點(diǎn),而關(guān)于肝內(nèi)膽管癌的研究卻相對(duì)較少,其發(fā)生發(fā)展機(jī)制尚不明確；丙酮酸激酶M2(PKM2)已被報(bào)道參與有氧和無氧糖酵解,并且已被證實(shí)和多種腫瘤發(fā)生和進(jìn)展相關(guān)。然而,在肝內(nèi)膽管癌組織中尚無關(guān)于PKM2的報(bào)道。本研究的目的是探討PKM2在肝內(nèi)膽管癌中的表達(dá)以及它肝內(nèi)膽管癌發(fā)生發(fā)展中的作用。 方法 收集50例肝內(nèi)膽管癌組織及其配對(duì)的癌旁組織,采用RT-PCR和免疫組織化學(xué)的方法檢測PKM2在肝內(nèi)膽管癌癌組織和配對(duì)癌旁組織中的表達(dá)水平,并分析其表達(dá)水平與肝內(nèi)膽管癌遠(yuǎn)期生存預(yù)后的關(guān)系；構(gòu)建PKM2的干擾RNA并轉(zhuǎn)染9810和CCLP-1細(xì)胞系,檢測PKM2蛋白的表達(dá)水平變化,并觀察對(duì)比9810和CCLP-1細(xì)胞侵襲、遷移和增殖能力的變化。 結(jié)果 RT-PCR結(jié)果提示肝內(nèi)膽管癌組織mRNA相對(duì)表達(dá)水平為3.24±0.693,顯著高于相對(duì)應(yīng)的癌旁組織的表達(dá)水平0.48±0.109,差異有統(tǒng)計(jì)學(xué)意義(P0.001)；免疫組化的結(jié)果表明:PKM2低表達(dá)組(評(píng)分≤7分)平均生存期為40.61±4.537月,明顯高于PKM2高表達(dá)組(評(píng)分7分)的15.15±2.368月,差異有統(tǒng)計(jì)學(xué)意義(P0.001)；在9810和CCLP-1細(xì)胞系中的,干擾組與對(duì)照組相比,在侵襲和轉(zhuǎn)移實(shí)驗(yàn)中,對(duì)照組細(xì)胞侵襲及遷移計(jì)數(shù)顯著高于干擾組(P0.001)；在增殖實(shí)驗(yàn)中隨著時(shí)間增長,在第48h、72h對(duì)照組的吸光度值顯著高于干擾組(P0.001)。 結(jié)論 PKM2在肝內(nèi)膽管癌組織中高表達(dá),并在肝內(nèi)膽管癌細(xì)胞的侵襲、遷移和增殖過程中起著重要作用,其表達(dá)程度高低還和肝內(nèi)膽管癌患者術(shù)后長期預(yù)后密切相關(guān)；PKM2可作為肝內(nèi)膽管癌診斷、靶向治療及判斷預(yù)后的潛在重要靶標(biāo)。
[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 鹵0.693, which was significantly higher than that in the corresponding paracancerous tissues (0.48 鹵0.109). The mean survival time was 40.61 鹵4.537 months. It was significantly higher than that in the high expression group of PKM2 (score 7) (15.15 鹵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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.8

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本文編號(hào)：1990471