Raf激酶抑制蛋白增强宫颈癌细胞的化疗敏感性的研究

发布时间：2018-04-15 17:53

  本文选题：Raf激酶抑制蛋白 + Hela细胞　； 参考：《青岛大学》2014年硕士论文

【摘要】：目的研究Raf激酶抑制蛋白对宫颈癌细胞化疗敏感性的影响。 方法以脂质体法将含有人全长RKIP基因的真核表达质粒pcDNA3.1(+)一ssRKIP转染入宫颈癌Hela细胞中,用G418筛选出含目的基因的稳定细胞系,Western blotting法检测Hela细胞中RKIP蛋白的表达。不同浓度顺铂作用不同时间,MTT法观察RKIP转染对顺铂作用于Hela细胞体外增值的影响,流式细胞术检测RKIP基因转染对顺铂诱导Hela细胞凋亡的影响。 结果pcDNA3.1(+)-ssRKIP转染的Hela细胞RKIP蛋白表达明显升高。不同浓度的顺铂分别处理各组细胞24、48、72h后,RKIP基因转染的Hela细胞增殖抑制率显著高于对照组细胞(P0.05)。以5μg/ml顺铂作用Hela细胞24h后,RKIP转染组细胞的凋亡率为(23.2±0.24)%,明显高于未转染组的(12.4±0.31)%和转染空质粒组的(13.4±0.47)%(P0.05)；在无顺铂作用情况下,RKIP转染组的细胞凋亡率为(5.7±0.12)%,仍然高于未转染组的(2.9±0.21)%和转染空质粒组的(3.6±0.08)%(P0.05)。 结论RKIP基因表达上调可以增强宫颈癌Hela细胞对化疗药物顺铂的敏感性。
[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 卤0.24, which was significantly higher than that in untransfected group (12.4 卤0.31%) and empty plasmid group (13.4 卤0.47P0.05%), and the apoptosis rate in RKIP transfection group was 5.7 卤0.12m without cisplatin, which was still higher than that in untransfected group.In group A, 2.9 卤0.21% in group A and 3.6 卤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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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