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目的 探讨沉默Notch1基因对人神经胶质瘤U251细胞增殖能力的影响.方法 采用Notch1-shRNA慢病毒感染人胶质瘤U251细胞,筛选稳定低表达Notch1的细胞单克隆,Western blot法检测细胞Notch1蛋白的表达;CCK-8法测定细胞生长;平板克隆形成实验和软琼脂克隆形成实验检测细胞的集落形成能力;裸鼠种植瘤模型观察Notch1沉默对种植瘤生长的影响.结果 成功筛选得到Notch1稳定沉默的U251细胞单克隆,其Notch1蛋白下调(65.3±5.1)%.Notch1稳定沉默细胞的生长增殖能力明显受到抑制,生长曲线与对照组相比明显减慢;Notch1沉默组的平板克隆形成率为(18.6±2.5)%,低于对照组(37.0±3.3)%;Notch1沉默组软琼脂集落形成率(51±5)%,低于对照组(80±4)%;Notch1沉默组裸鼠腋下种植瘤生长速度减慢,30 d后瘤重(0.31±0.09)g,明显低于对照组(2.35±0.71)g;统计学分析差异均有显著性.结论 RNAi沉默Notch1基因可致人胶质瘤U251细胞的体外增殖和体内成瘤能力下降.“,”Aim To investigate the effects of Notch1 knockdown by RNA interference(RNAi)on proliferation of human glioma U251 cells.Methods The human glioma U251 cells were infected with lentivirus expressing Notch1-shRNA.The knockdown effect of Notch1 in transduced U251 cells was detected by Western blot.The cell growth viability was evaluated by CCK-8 assay.Colony formation assay and soft-agar colony formation were used to measure the colony formation ability of stably transduced cells.The influence of Notch1-RNAi on the implanted tumor growth was observed.Results The Notch1-knockdown U251 cell clone was selected.Notch1 expression was down-regulated by Notch1-shRNA, and the inhibitory rate was(65.3±5.1)%.The ability of growth in Notch1-knockdown cells was decreased significantly.The plate colony formation rate of Notch1-knockdown cells was(18.6±2.5)%,whereas the rate of control cells was(37.0±3.3)%.The soft-agar colony formation rate of Notch1-knockdown cells was(51±5)%,whereas the rate of control cells was (80±4)%.Implanted glioma mouse model was successfully established.The tumor weight in Notch1-knockdown group was markedly lower than that in control group(0.31±0.09 g vs 2.35±0.71g,P<0.01).Conclusion Down-regulation of Notch1 expression by RNAi can inhibit the proliferation of human glioma U251 cells.