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医学论文范文:RNA干扰抑制Survivin表达对人喉癌细胞药物敏感性的影响
【摘要】 目的：探讨RNA干扰技术抑制Survivin基因表达对增强人喉癌HEP2细胞化疗药物敏感性。方法:针对Survivin mRNA序列设计合成小干扰RNA(siRNA)的DNA模板,构建pRNATSurvivin重组表达载体，转染人喉癌HEP2细胞;通过RTPCR和Western blot分析其对HEP2细胞内源性Survivin表达的影响;四甲基偶氮唑蓝(MTT)法观察HEP2细胞对化疗药物5FU和优福定敏感性的改变。结果:成功构建pRNATSurvivin重组质粒,并成功转染HEP2细胞;RTPCR和Western blot结果证实重组质粒在mRNA及蛋白水平分别显著抑制Survivin基因表达(P<0.01);MTT及流式细胞仪结果证明在5FU和优福定联合作用下，pRNATSurvivin组HEP2细胞的生长抑制率明显增高(P<0.01)。结论:pRNATSurvivin可抑制Survivin在人喉癌HEP2细胞中的表达,并增强HEP2细胞对化疗药物敏感性。
【关键词】 Survivin RNA干扰 喉癌 药物敏感性
Effect of Inhibited Expression of Survivin by RNA Interference on the Drug Sensitivity in Human Laryngocarcinoma Cells CHEN Xiangjun,YU Zizhong,LI Guoyi^ (Department of Otolaryngology,Taihe Hospital,Yunyang Medical College,Shiyan,Hubei 442000,China)
Abstract:Objective To explore the effect of inhibited expression of survivin by RNA interference on sensitivity of HEP2 cells to chemotherapeutic drugs. Methods One pair of DNA template coding siRNA was designed and synthesized against Survivin mRNA sequence,the pRNATSurvivin recombinant expression vector was constructed and transfected into HEP2 cells.The effect of endogenous survivin expression in HEP2 cells by RNA interference was analyzed with RTPCR and Western blot.The changes of sensitivity to 5FU and UFT in HEP2 cells were observed with MTT.Results Recombinant plasmid pRNATSurvivin was constructed and transfected into HEP2 cells successfully,recombinant plasmid pRNATSurvivin could significantly inhibit the expression of Survivin in HEP2 cells (P<0.01),which was demonstrated by RTPCR and Western blot.The growth inhibitory rate of HEP2 cells in pRNATSurvivin group was significantly increased under the combined effect of 5FU and UFT(P<0.01),which was demonstrated by MTT and flow cytometry. Conclusion pRNAT Survivin could inhibit the expression of Survivin,enhance the sensitivity of HEP2 cells to chemotherapeutic drugs医.学全.在.线网站www.lindalemus.com.
Key words: Survivin;RNA interference;Laryngocarcinoma;Sensitivity
