摘要
目的观察靶向survivin基因的短发夹RNA(shorthairpin RNA,shRNA)对人肺腺癌细胞A549生物学行为及紫杉醇敏感性的影响。方法将靶向survivin的基因片段插入载体后构建重组质粒,用转染试剂FuGENE(HD将其导入A549细胞,RT-PCR及Western blot分析转染前后survivin基因的表达情况,TUNEL法检测细胞凋亡情况,MTT法检测转染后A549细胞对紫杉醇的敏感性变化。结果成功构建重组质粒。与对照组相比,转染重组质粒后,survivin基因的表达明显降低,细胞凋亡率增加。转染前紫杉醇抑制A549细胞的IC50为转染后的11.9倍,两者比较,差异有显著性(P<0.05)。结论靶向survivin的shRNA能下调survivin基因表达,诱导细胞凋亡,增强A549细胞对紫杉醇的敏感性。
Aim To construct the eukaryotic expression vectors of short hairpin RNA targeting survivin and observe its effect on biologic behavior of A549 cells and sensitivity of A549 cells to paclitaxel. Methods The DNA fragment targeting human survivin was inserted into the plasmid, and the recombinant plasmid was constructed. The recombinant plasmids cells were transfected into A549 cells by FuGENE transfection reagent. The expression levels of survivin gene were detected with RT-PCR and Western blot before and after transfection, respectively. Cell apoptosis was detected by TUNEL method. The sensitivity of A549 cells to paclitaxel was detected by MTT after transfection. Results The recombinant plasmid was successfully constructed. RNAi group cells showed lower expression of survivin than control group. The apoptosis rate of A549 cells increased after transfection. The IC50 of paclitaxel inhibiting A549 cells was 11.9 fold before transfection compared with those after transfection. There was significant difference between the two groups （P 〈 0. 05 ）. Conclusion The shRNA targeting human survivin can down-regulate the expression of survivin, induce apoptosis, and enhance the sensitivity of A549 cells to paclitaxel.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2009年第5期589-592,共4页
Chinese Pharmacological Bulletin
基金
山东省自然科学基金青年基金资助项目(NoQ2006D08)
