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目的探讨Survivin短发卡状RNA抑制Survivin的表达对人乳腺癌MCF-7细胞紫杉醇敏感性的影响。方法通过脂质体介导将Survivin短发卡RNA表达质粒pSurvivin shRNA转染人乳腺癌细胞系MCF-7并检测Survivin mRNA及蛋白的表达情况;噻唑蓝(MTF)比色法和An- nexin-V法分别检测紫杉醇处理后转染pSurvivin shRNA细胞、未转染细胞以及转染阴性对照质粒细胞的增殖和细胞凋亡的变化;采用Western blot法观察紫杉醇作用过程中Survivin表达的变化。结果与未转染细胞及转染阴性对照质粒细胞比较,转染pSurvivitt shRNA质粒的细胞Survivin mRNA和蛋白表达明显下降;在同一紫杉醇作用浓度下,细胞增殖抑制率和细胞凋亡率明显增高。此外在紫杉醇作用的早期(6h内),转染阴性对照质粒细胞以及未转染细胞均出现了Survivin表达一过性增加,而转染pSurvivin shRNA的细胞中未观察到Survivin表达的增加。结论短发卡状RNA干扰技术阻抑乳腺癌细胞中Survivin的表达,可增强乳腺癌细胞对紫杉醇的敏感性。
Objective To investigate the effect of Survivin short hairpin RNA on the paclitaxel sensitivity of human breast cancer MCF-7 cells induced by Survivin. Methods Survivin short hairpin RNA expression plasmid pSurvivin shRNA was transfected into human breast cancer cell line MCF-7 by liposome and the expression of Survivin mRNA and protein was detected by MTT assay and An-nexin-V Method were used to detect paclitaxel-treated pSurviv shRNA transfected cells, untransfected cells and transfected negative control plasmid cell proliferation and apoptosis were observed by Western blot assay paclitaxel during Survivin expression changes. Results Compared with untransfected cells and transfected negative control plasmid, Survivin mRNA and protein expression in transfected pSurvivitt shRNA plasmid was significantly decreased. Under the same paclitaxel concentration, the cell proliferation inhibition rate and apoptosis rate were significantly increased. In addition, Survivin expression was transiently increased in transfected negative control plasmid cells and untransfected cells in early (within 6h) paclitaxel treatment, whereas no increase in Survivin expression was observed in cells transfected with pSurvivin shRNA. Conclusion Short hairpin RNA interference technology inhibits the expression of Survivin in breast cancer cells and enhances the sensitivity of breast cancer cells to paclitaxel.