论文部分内容阅读
目的:采用RNA干扰技术下调Notch2基因在Lewis肺癌细胞(LLC)的表达,探讨Notch2表达下调对LLC增殖和周期的影响。方法:通过脂质体2000将Notch2-siRNA和对照con-siRNA分别转染LLC后,通过PCR,RT-PCR检测Notch2的表达情况;通过MTT、软琼脂克隆形成实验检测LLC的生长情况;并使用流式细胞技术检测细胞周期。结果:SiRNA转染LLC48小时后,PCR和RT-PCR检测结果显示Notch2在实验组的表达受到显著抑制;MTT实验显示干扰Notch2的表达显著抑制了LLC的增殖(P<0.001);软琼脂克隆形成实验表明干扰Notch2的表达将影响LLC的克隆形成;细胞周期实验进一步证实干扰Notch2的表达使处于活跃增殖期(S期)的LLC数量明显减少。结论:Notch2在LLC中的表达发挥重要的促癌作用,干扰Notch2的表达能显著抑制LLC的分裂增殖和克隆形成,从而抑制肿瘤的生长。
OBJECTIVE: To investigate the effect of down-regulation of Notch2 on the proliferation and the cycle of LLC by down-regulating the expression of Notch2 gene in Lewis lung carcinoma (LLC) cells by RNAi technique. METHODS: Notch2-siRNA and control con-siRNA were transfected into LLC by Lipofectamine 2000 respectively. The expression of Notch2 was detected by PCR and RT-PCR. The growth of LLC was detected by MTT and soft agar colony formation assay. Flow Cytometry detects cell cycle. Results: After48h siRNA transfection, the expression of Notch2 in the experimental group was significantly inhibited by PCR and RT-PCR. The MTT assay showed that the expression of Notch2inhibited the proliferation of LLC significantly (P <0.001) Experiments show that the interference of Notch2 expression will affect the formation of LLC clones; cell cycle experiments further confirmed that the interference of Notch2 expression in the active proliferative phase (S phase) significantly reduced the number of LLC. Conclusion: The expression of Notch2 in LLC plays an important role in carcinogenesis. Interference of Notch2 expression significantly inhibits the growth and proliferation of LLC cells.