RNAi对GBC-SD细胞survivin基因表达及阿霉素敏感性的影响

摘要

目的 利用RNA干扰(KNAi)技术沉默人胆囊癌GBC-SD细胞中抗凋亡基因survivin,观察survivin基因表达的变化以及其对阿霉素的敏感性.方法 将靶向survivin的基因片段插入到栽体后构建重组质粒,将其导入GBC-SD细胞,RT-PCR及Westem-blotting法检测转染前后GBC-SD细胞survivin mRNA及蛋白水平的表达情况,MTT法检测转染前后GBC-SD细胞对阿霉素的敏感性变化.结果 成功构建重组质粒.转染重组质粒后,GBC-SD细胞survivin mRNA和蛋白的表达明显降低;转染前后阿霉素对GBC-SD细胞生存率具显著影响.结论 靶向survivin的RNAi表达载体能下调survivin基因表达,进而增强阿霉素对GBC-SD细胞化疗敏感性.%Objective To use RNA interference (RNAi) technology to silence anti apoptosis gene survivin of human gallbladder carcinoma GBC-SD cells, and then to observe changes of survivin expressions and the sensitivity to doxorubicin. Methods Targeting survivin gene fragment was inserted into the carrier, and then the constructed recombinant plasmids were induced to GBC-SD cells. The mRNA and protein levels of survivin were detected with RT-PCR and Western-blotting respectively before and after transfection. The sensitivity to doxorubicin was detected with MTT before and after transfection. Results The recombinant plasmids were successfully constructed. After transfection of recombinant plasmid in GBC-SD cells, the mRNA and protein expressions of survivin were significantly reduced; meanwhile doxorubicin could significantly inhibit GBC-SD cell growth. Conclusion The RNA interference expression vectors targeting surviving, which down-regulate the expression of surviving, can enhance the sensitivity of GBC-SD cells to doxorubicin.