RNA干扰抑制Survivin表达对人喉癌细胞药物敏感性的影响

目的:探讨RNA干扰技术抑制Survivin基因表达对增强人喉癌HEP-2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pRNAT-Survivin重组表达载体,转染人喉癌HEP-2细胞;通过RT-PCR和Western blot分析其对HEP-2细胞内源性Survivin表达的影响;四甲基偶氮唑蓝(MTT)法观察HEP-2细胞对化疗药物5-FU和优福定敏感性的改变。结果:成功构建pRNAT-Survivin重组质粒,并成功转染HEP-2细胞;RT-PCR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5-FU和优福定联合作用下,pRNAT-Survivin组HEP-2细胞的生长抑制率明显增高(P<0.01)。结论:pRNAT-Survivin可抑制Survivin在人喉癌HEP-2细胞中的表达,并增强HEP-2细胞对化疗药物敏感性。

Effect of Inhibited Expression of Survivin by RNA Interference on the Drug Sensitivity in Human Laryngocarcinoma Cells

Objective To explore the effect of inhibited expression of survivin by RNA interference on sensitivity of HEP-2 cells to chemotherapeutic drugs. Methods One pair of DNA template coding siRNA was designed and synthesized against Survivin mRNA sequence,the pRNAT-Survivin recombinant expression vector was constructed and transfected into HEP-2 cells. The effect of endogenous survivin expression in HEP-2 cells by RNA interference was analyzed with RT-PCR and Western blot. The changes of sensitivity to 5-FU and UFT in HEP-2 cells were observed with MTI＇. Results Recombinant plasmid pRNAT-Survivin was constructed and transfected into HEP-2 cells successfully, recombinant plasmid pRNAT-Survivin could significantly inhibit the expression of Survivin in HEP-2 cells （P 〈 0.01 ） ,which was demonstrated by RT-PCR and Western blot. The growth inhibitory rate of HEP-2 cells in pRNAT-Survivin group was significandy increased under the combined effect of 5-FU and FT（ P 〈0.01 ） ,which was demonstrated by MTT and flow eytometry. Conclusion pRNAT-Survivin could inhibit the expression of Survivin, enhance the sensitivity of HEP-2 cells to chemotherapeutic drugs.