【作者】 吴雨田；

【导师】 陈燕凌；

【作者基本信息】 福建医科大学 ， 外科学， 2009， 硕士

【摘要】 目的:探讨幽门螺杆菌(Hp)对人类胆管癌细胞增殖和凋亡的影响,以及该影响的细胞内信号转导途径。方法:用CCK-8(Cell Counting Kit-8)法和流式细胞仪法检测Hp对胆管癌细胞增殖,凋亡及细胞周期分布情况的影响,利用免疫细胞化学和凝胶电泳迁移率变动实验(EMSA)分析细胞核转录因子NF-κB在Hp作用前后的表达活化情况。再使用特异性NF-κB阻断剂吡咯烷二硫氨基甲酸酯(PDTC),阻断NF-κB的活化,利用细胞免疫化学技术和RT-PCR半定量技术分析比较抗凋亡蛋白Bcl-2和Bcl-XL在转录因子NF-κB失活前后的表达情况。结果:在MOI值(细菌与细胞浓度比)为0.5:1的状态下, Hp对QBC939细胞有促增殖作用,第1,3天的增殖率分别为15.24%±4.2%、34.18%±2.6%,t检验P<0.05,具有统计学意义。在该MOI值时无诱导细胞凋亡作用,但可推动细胞周期的进展;在此过程中伴有细胞NF-κB活化和抗凋亡蛋白Bcl-2,Bcl-XL表达上调;PDTC阻断实验证明抗凋亡蛋白Bcl-2,Bcl-XL的表达受到NF-κB活性的调控。结论:幽门螺杆菌—核转录因子NF-κB—抗凋亡蛋白Bcl-2,Bcl-XL,这一传导通路的活化,在胆管癌细胞的增殖,凋亡及恶性转化中很可能具有重要作用。更多还原

【Abstract】 Purpose: To discuss the influence on proliferation and apoptosis of human bile duct carcinoma cell by Helicobacter pylori (Hp); and signal transduction pathways in cells of this influence.Methods: Apply the CCK-8 (Cell Counting Kit-8) method and flow cytometry to test the influence on proliferation and apoptosis of bile duct carcinoma cell by Helicobacter pylori and cell cycle distribution, utilize the immunocytochemistry and gel electrophoretic mobility shift assay ( EMSA)to analyze the expression and activation conditions of nuclear transcription factor NF-KB before and after Hp. And then use the specific NF-KB inhibitor pyrrolidine dithiocarbamate (PDTC) to inhibit the activation of NF-KB, and utilize the cellular immunity chemical technology and RT-PCR semi quantitative to analyze and compare the expression conditions of anti-apoptosis protein Bcl-2 and Bcl-XL before and after deactivation of NF-KB.Result: At MOI (concentration ratio of bacteria to cell) of 0.5:1, Hp could facilitate the proliferation of QBC939 cells, the breeding ratios on the first and third day are 15.24%±4.2% and 34.18%±2.6% respectively, t test, P<0.05, having statistical significance. There is no induced Apoptosis at this MOI value, but could promote the development of cell cycle; the cell NF-KB is activated and the expression of anti-apoptosis protein Bcl-2 and Bcl-XL is up-regulated; the PDTC inhibition test proves that the expression of anti-apoptosis protein Bcl-2 and Bcl-XL is controlled by the activity of NF-KB.Conclusion: Helicobacter pylori--nuclear factor NF-KB--anti-apoptosis protein Bcl-2, Bcl-XL, the activation of this pathway is likely to play a very important role in the proliferation, apoptosis and malignant transformation of bile duct carcinoma cell.更多还原