本文關鍵詞：TRAIL在HepG2細胞表達及細胞凋亡機制的研究　出處：《青島大學》2015年碩士論文　論文類型：學位論文

  更多相關文章： HepG2 TRAIL 慢病毒載體 細胞凋亡

【摘要】：目的:把攜帶TRAIL基因的慢病毒載體構建在Hep G2細胞中表達,檢測TRAIL對Hep G2的蛋白表達及增殖,觀察TRAIL-Hep G2細胞與化療藥物聯(lián)用效果,并探討其可能凋亡機制方法:1、肝癌細胞株Hep G2細胞培養(yǎng)。2、構建TRAIL重組慢病毒表達載體p CDH-CMV-TRAIL-EF1-GFP-T2A-Puro。3、慢病毒感染肝癌細胞Hep G2的TRAIL蛋白表達3、化療藥物干預TRAIL-Hep G2細胞。4、MTT檢測細胞抑制率。5、流式細胞術檢測細胞周期。結果:測序證實成功構建攜帶TRAIL基因慢病毒載體,當MOI=40時,重組慢病毒體在Hep G2細胞的外轉染效率高表達。經Western blotting方法檢測表達的TRAIL蛋白。MTT檢測發(fā)現(xiàn)轉染TRAIL-Hep G2藥物組細胞對細胞增殖活力的影響明顯低于Hep G2和TRAIL-Hep G2組(P0.05),流式細胞學檢對Hep G2細胞周期影響,得出凋亡比例中實驗組明顯高于對照組,且p值0.05,有統(tǒng)計學意義意義。結論:首先成功構建了帶有TRAIL基因的慢病毒載體,并在Hep G2的穩(wěn)定高表達。通過在實驗組及對照組中應用濃度為0.1μg/m L化療藥物5-FU,兩組的凋亡比例具有明顯的差別,統(tǒng)計學有意義(P0.05)。說明該藥物與TRAIL介導的Hep G2細胞協(xié)同作用能加速凋亡過程,并能明顯抑制肝癌細胞的生長與繁殖。
[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.

【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.7

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本文編號：1359233