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醫(yī)學論文范文:RNA干擾抑制Survivin表達對人喉癌細胞藥物敏感性的影響
【摘要】 目的：探討RNA干擾技術抑制Survivin基因表達對增強人喉癌HEP2細胞化療藥物敏感性。方法:針對Survivin mRNA序列設計合成小干擾RNA(siRNA)的DNA模板,構建pRNATSurvivin重組表達載體，轉染人喉癌HEP2細胞;通過RTPCR和Western blot分析其對HEP2細胞內源性Survivin表達的影響;四甲基偶氮唑藍(MTT)法觀察HEP2細胞對化療藥物5FU和優(yōu)福定敏感性的改變。結果:成功構建pRNATSurvivin重組質粒,并成功轉染HEP2細胞;RTPCR和Western blot結果證實重組質粒在mRNA及蛋白水平分別顯著抑制Survivin基因表達(P<0.01);MTT及流式細胞儀結果證明在5FU和優(yōu)福定聯(lián)合作用下，pRNATSurvivin組HEP2細胞的生長抑制率明顯增高(P<0.01)。結論:pRNATSurvivin可抑制Survivin在人喉癌HEP2細胞中的表達,并增強HEP2細胞對化療藥物敏感性。
【關鍵詞】 Survivin RNA干擾 喉癌 藥物敏感性
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醫(yī).學全.在.線網(wǎng)站www.gydjdsj.org.cn.
Key words: Survivin;RNA interference;Laryngocarcinoma;Sensitivity
