本文關(guān)鍵詞：TRAIL在HepG2細(xì)胞表達(dá)及細(xì)胞凋亡機(jī)制的研究　出處：《青島大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： HepG2 TRAIL 慢病毒載體 細(xì)胞凋亡

【摘要】：目的:把攜帶TRAIL基因的慢病毒載體構(gòu)建在Hep G2細(xì)胞中表達(dá),檢測TRAIL對Hep G2的蛋白表達(dá)及增殖,觀察TRAIL-Hep G2細(xì)胞與化療藥物聯(lián)用效果,并探討其可能凋亡機(jī)制方法:1、肝癌細(xì)胞株Hep G2細(xì)胞培養(yǎng)。2、構(gòu)建TRAIL重組慢病毒表達(dá)載體p CDH-CMV-TRAIL-EF1-GFP-T2A-Puro。3、慢病毒感染肝癌細(xì)胞Hep G2的TRAIL蛋白表達(dá)3、化療藥物干預(yù)TRAIL-Hep G2細(xì)胞。4、MTT檢測細(xì)胞抑制率。5、流式細(xì)胞術(shù)檢測細(xì)胞周期。結(jié)果:測序證實成功構(gòu)建攜帶TRAIL基因慢病毒載體,當(dāng)MOI=40時,重組慢病毒體在Hep G2細(xì)胞的外轉(zhuǎn)染效率高表達(dá)。經(jīng)Western blotting方法檢測表達(dá)的TRAIL蛋白。MTT檢測發(fā)現(xiàn)轉(zhuǎn)染TRAIL-Hep G2藥物組細(xì)胞對細(xì)胞增殖活力的影響明顯低于Hep G2和TRAIL-Hep G2組(P0.05),流式細(xì)胞學(xué)檢對Hep G2細(xì)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(xué)意義意義。結(jié)論:首先成功構(gòu)建了帶有TRAIL基因的慢病毒載體,并在Hep G2的穩(wěn)定高表達(dá)。通過在實驗組及對照組中應(yīng)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(xué)有意義(P0.05)。說明該藥物與TRAIL介導(dǎo)的Hep G2細(xì)胞協(xié)同作用能加速凋亡過程,并能明顯抑制肝癌細(xì)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(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R735.7

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本文編號：1359233