Objective: To understand the function of adaptor protein p66She, one approach of silencing p66She through shRNA lentivirus in non-small cell lung cancer cell A549. Methods:Semi RT-PCR and Western blot were used to investigate P66She knockdown effect at the mRNA and protein levels, and the proliferation change was detected by BrdU labeling assay. Results:In AS49 cells, mRNA and protein level of p66She were reduced significantly by its specific shRNA interference. Also, AS49 cells proliferation was dramatically attenuated after p66She knockdown. Conclusion:Highly transfective effect is acquired in AS49 by lentivirus mediated p66She-shRNA, resulting in specific gene silencing effect, and significant inhibition of cell proliferation. p66She-specific gene knockdown may be a useful experimental method for investigating p66She function.%目的:在人非小细胞肺癌细胞A549细胞中探讨shRNA靶向慢病毒对信号衔接蛋白p66Shc的沉默效应,以更好地研究p66Shc的功能.方法:半定量RT- PCR和Western blot检测p66Shc下调后的变化情况,BrdU标记实验检测细胞的增殖变化情况.结果:在A549细胞中,运用p66Shc-shRNA干扰可以实现p66Shc在mRNA和蛋白水平的下调作用,同时发现A549细胞的增殖显著降低.结论:经过慢病毒栽体介导的p66Shc-shRNA在人非小细胞肺癌A549中可获得高效转染效果,并能产生特异性的基因沉默效应,而且对细胞的增殖有显著的抑制作用,为进一步研究p66Shc功能提供了实验手段.
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