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背景与目的:survivin属IAP基因家族成员,表达于多种肿瘤组织中,能促使细胞逃避凋亡,并促进细胞的异常有丝分裂。本研究旨在探讨敲除survivin基因后,癌细胞的增殖和凋亡情况。方法:利用RNAi阻抑人乳腺癌细胞内survivin基因的表达,RT-PCR和Westernblot法分析survivin基因mRNA和蛋白的表达,MTT法检测细胞生长增殖抑制率,流式细胞术检测细胞凋亡率。结果:survivinsiRNA转染组,survivin基因表达水平与未转染组比较下调了64%。随着siRNA浓度的增加,细胞增殖抑制率逐渐增高,200nmol/L剂量组细胞增殖抑制率最高,可达60.9%。不同浓度的siRNA可不同程度地诱导细胞凋亡,200nmol/L剂量组凋亡率最高,可达29.0%。结论:survivinsiRNA有可能成为治疗乳腺癌的新药物。
BACKGROUND & OBJECTIVE: Survivin is a member of the IAP gene family and is expressed in many tumor tissues. It can promote apoptosis of cells and promote abnormal mitosis. The aim of this study was to investigate the proliferation and apoptosis of cancer cells after the knockdown of survivin gene. Methods: The expression of survivin gene in human breast cancer cells was inhibited by RNAi. The expression of survivin mRNA and protein was analyzed by RT-PCR and Western blotting. The inhibition rate of cell growth was detected by MTT assay. The apoptosis rate was detected by flow cytometry. Results: The survivin gene expression level was down-regulated by 64% in survivin siRNA transfection group. With the increase of siRNA concentration, the cell proliferation inhibition rate gradually increased, 200nmol / L dose group, the highest inhibition rate of cell proliferation up to 60.9%. Different concentrations of siRNA induced apoptosis to different extents, and the highest apoptosis rate was reached at 200 nmol / L (29.0%). Conclusion: survivinsiRNA may become a new drug for the treatment of breast cancer.