TRAIL在HepG2细胞表达及细胞凋亡机制的研究

发布时间：2017-12-31 09:40

本文关键词：TRAIL在HepG2细胞表达及细胞凋亡机制的研究　出处：《青岛大学》2015年硕士论文　论文类型：学位论文

更多相关文章： HepG2 TRAIL 慢病毒载体 细胞凋亡

【摘要】：目的:把携带TRAIL基因的慢病毒载体构建在Hep G2细胞中表达,检测TRAIL对Hep G2的蛋白表达及增殖,观察TRAIL-Hep G2细胞与化疗药物联用效果,并探讨其可能凋亡机制方法:1、肝癌细胞株Hep G2细胞培养。2、构建TRAIL重组慢病毒表达载体p CDH-CMV-TRAIL-EF1-GFP-T2A-Puro。3、慢病毒感染肝癌细胞Hep G2的TRAIL蛋白表达3、化疗药物干预TRAIL-Hep G2细胞。4、MTT检测细胞抑制率。5、流式细胞术检测细胞周期。结果:测序证实成功构建携带TRAIL基因慢病毒载体,当MOI=40时,重组慢病毒体在Hep G2细胞的外转染效率高表达。经Western blotting方法检测表达的TRAIL蛋白。MTT检测发现转染TRAIL-Hep G2药物组细胞对细胞增殖活力的影响明显低于Hep G2和TRAIL-Hep G2组(P0.05),流式细胞学检对Hep G2细胞周期影响,得出凋亡比例中实验组明显高于对照组,且p值0.05,有统计学意义意义。结论:首先成功构建了带有TRAIL基因的慢病毒载体,并在Hep G2的稳定高表达。通过在实验组及对照组中应用浓度为0.1μg/m L化疗药物5-FU,两组的凋亡比例具有明显的差别,统计学有意义(P0.05)。说明该药物与TRAIL介导的Hep G2细胞协同作用能加速凋亡过程,并能明显抑制肝癌细胞的生长与繁殖。
[Abstract]:Objective: to construct a lentiviral vector carrying TRAIL gene expression in Hep G2 cells, protein expression and proliferation of Hep G2 detection of TRAIL, TRAIL-Hep and G2 cells were combined with chemotherapy, and to investigate its possible mechanism. Methods: 1, apoptosis of hepatocellular carcinoma cell line Hep G2 cell culture.2, construction of recombinant TRAIL slow virus expression vector p CDH-CMV-TRAIL-EF1-GFP-T2A-Puro.3 3 expression lentivirus infected hepatoma cells Hep G2 TRAIL protein, TRAIL-Hep chemotherapy G2 cell.4, MTT detection of cell inhibition rate of.5 cell cycle by flow cytometry. Results: sequencing successfully constructed lentiviral vector carrying TRAIL gene, when MOI=40, the recombinant lentivirus infected body the high expression efficiency in Hep G2 cells. TRAIL protein. Detection of.MTT expression detected by Western blotting method found the effect of transfection of TRAIL-Hep G2 drug group cells on cell proliferation activity of the Ming Dynasty Hep G2 and TRAIL-Hep were lower than group G2 (P0.05), flow cytometry detection effect on Hep G2 cell cycle, the percentage of apoptosis in experimental group was significantly higher than the control group, and the p value is 0.05, there was statistical significance. Conclusion: first successfully constructed lentiviral vector carrying TRAIL gene, and Hep in stable G2 high expression. Through the application in experiment group and control group was 0.1 g/m L 5-FU chemotherapy, the apoptosis rate of the two groups have obvious differences, statistically significant (P0.05). The drug and TRAIL mediated Hep G2 cell can accelerate the apoptosis process of synergistic effect, and can significantly inhibit the growth and the breeding of hepatocellular carcinoma cells.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.7

【参考文献】