本文選題：Raf激酶抑制蛋白 + Hela細(xì)胞　； 參考：《青島大學(xué)》2014年碩士論文

【摘要】：目的研究Raf激酶抑制蛋白對(duì)宮頸癌細(xì)胞化療敏感性的影響。 方法以脂質(zhì)體法將含有人全長(zhǎng)RKIP基因的真核表達(dá)質(zhì)粒pcDNA3.1(+)一ssRKIP轉(zhuǎn)染入宮頸癌Hela細(xì)胞中,用G418篩選出含目的基因的穩(wěn)定細(xì)胞系,Western blotting法檢測(cè)Hela細(xì)胞中RKIP蛋白的表達(dá)。不同濃度順鉑作用不同時(shí)間,MTT法觀察RKIP轉(zhuǎn)染對(duì)順鉑作用于Hela細(xì)胞體外增值的影響,流式細(xì)胞術(shù)檢測(cè)RKIP基因轉(zhuǎn)染對(duì)順鉑誘導(dǎo)Hela細(xì)胞凋亡的影響。 結(jié)果pcDNA3.1(+)-ssRKIP轉(zhuǎn)染的Hela細(xì)胞RKIP蛋白表達(dá)明顯升高。不同濃度的順鉑分別處理各組細(xì)胞24、48、72h后,RKIP基因轉(zhuǎn)染的Hela細(xì)胞增殖抑制率顯著高于對(duì)照組細(xì)胞(P0.05)。以5μg/ml順鉑作用Hela細(xì)胞24h后,RKIP轉(zhuǎn)染組細(xì)胞的凋亡率為(23.2±0.24)%,明顯高于未轉(zhuǎn)染組的(12.4±0.31)%和轉(zhuǎn)染空質(zhì)粒組的(13.4±0.47)%(P0.05)；在無順鉑作用情況下,RKIP轉(zhuǎn)染組的細(xì)胞凋亡率為(5.7±0.12)%,仍然高于未轉(zhuǎn)染組的(2.9±0.21)%和轉(zhuǎn)染空質(zhì)粒組的(3.6±0.08)%(P0.05)。 結(jié)論RKIP基因表達(dá)上調(diào)可以增強(qiáng)宮頸癌Hela細(xì)胞對(duì)化療藥物順鉑的敏感性。
[Abstract]:Objective to study the effect of Raf kinase inhibitor protein on chemosensitivity of cervical cancer cells.Methods the eukaryotic expression plasmid pcDNA3.1 (ssRKIP) containing human full-length RKIP gene was transfected into cervical cancer Hela cells by liposome method. The stable cell line containing the target gene was screened by G418 and the expression of RKIP protein in Hela cells was detected by Western blotting.The effects of RKIP transfection on the proliferation of Hela cells induced by cisplatin in vitro were observed by MTT assay. The effect of RKIP gene transfection on the apoptosis of Hela cells induced by cisplatin was detected by flow cytometry.Results the expression of RKIP protein was significantly increased in Hela cells transfected with pcDNA3.1 (-ssRKIP).The proliferation inhibition rate of Hela cells transfected with RKIP gene was significantly higher than that of control cells treated with different concentrations of cisplatin for 72 h.After treated with 5 渭 g/ml cisplatin for 24 hours, the apoptotic rate of Hela cells in RKIP transfection group was 23.2 鹵0.24, which was significantly higher than that in untransfected group (12.4 鹵0.31%) and empty plasmid group (13.4 鹵0.47P0.05%), and the apoptosis rate in RKIP transfection group was 5.7 鹵0.12m without cisplatin, which was still higher than that in untransfected group.In group A, 2.9 鹵0.21% in group A and 3.6 鹵0.08 in group A in empty plasmid transfection group (P 0.05).Conclusion upregulation of RKIP gene expression can enhance the sensitivity of cervical cancer Hela cells to cisplatin.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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