【摘要】 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:经过慢病毒载体介导的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。更多还原

【Abstract】 Objective: To understand the function of adaptor protein p66 Shc,one approach of silencing p66 Shc through shRNA lentivirus in non-small cell lung cancer cell A549.Methods : Semi RT-PCR and Western blot were used to investigate p66 Shc knockdown effect at the mRNA and protein levels,and the proliferation change was detected by BrdU labeling assay.Results : In A549 cells,mRNA and protein level of p66 Shc were reduced significantly by its specific shRNA interference.Also,A549 cells proliferation was dramatically attenuated after p66 Shc knockdown.Conclusion : Highly transfective effect is acquired in A549 by lentivirus mediated p66 Shc-shRNA,resulting in specific gene silencing effect,and significant inhibition of cell proliferation.p66 Shc-specific gene knockdown may be a useful experimental method for investigating p66 Shc function.更多还原