摘要
目的探讨siRNA沉默Msi1基因表达对人结肠癌SW-480细胞增殖的影响。方法针对Msi1 mRNA序列设计合成siRNA,转染SW-480细胞;采用RT-PCR法检测Msi1基因表达;MTT比色实验、群体倍增时间及平皿克隆形成实验分析Msi1siRNA对人结肠癌SW-480细胞的生长抑制效应,流式细胞术检测Msi1 siRNA对其细胞周期的影响。结果 Msi1 siRNA能有效抑制人结肠癌SW-480细胞中Msi1基因的表达;Msi1 siRNA转染SW-480细胞24、48、72 h后,与其它各组比较,其吸光度值明显降低,Msi1 siRNA组SW-480细胞生长、增殖速度明显减缓(P<0.05);空白对照组的SW-480细胞群体倍增时间为22.15h,而Msi1 siRNA处理后的细胞,群体倍增时间延长至37.76 h(P<0.05);平皿克隆形成实验结果发现,Msi1 siRNA处理后的细胞与其它各组相比,细胞生长明显减慢。流式细胞术表明Msi1 siRNA可导致SW-480细胞G0/G1期阻滞,S期细胞减少。结论沉默Msi1基因对人结肠癌SW-480细胞增殖有负性调节作用。
Purpose To investigate the effect of silencing Msi1 gene with siRNA on cell proliferation of SW-480 cell line.Methods Msi1-specific siRNA was designed and synthesized,and the small interfering RNA was transfected into SW-480 cells to knockdown the Msi1 expression.mRNA levels of Msi1 was detected by RT-PCR,MTT assay,cell average doubling time analysis and plate cloning experiment were employed to confirm Msi1 siRNA-induced cell growth inhibition in cell line SW-480.Results RNA interference efficiently suppressed the Msi1 expression in SW-480 cell line.After SW-480 cells transfected with Msi1 siRNA with 24 h,48 h and 72 h,proliferation rate was reduced（P0.05）;average doubling time retarded from 22.15 hours in normal cultured SW-480 cells to 37.76 hours in Msi1 siRNA experimented SW-480 cells（P0.05）.Plate cloning experiment showed the growth rate of Msi1 siRNA group was lower than that in other groups.Conclusions Downregulation of Msi1 results in significant inhibition of tumor growth in the SW-480 cell line.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2012年第2期124-127,共4页
Chinese Journal of Clinical and Experimental Pathology
基金
浙江省医学重点学科建设计划(GJSX-010-004)
