禽流感病毒NS1A蛋白诱导人肺癌细胞SPC-A1的凋亡

目的研究禽流感病毒NS1A蛋白对人肺癌细胞SPC-A1增殖的影响。方法将含有NS1A基因的重组质粒pVAX1-NS1A经脂质体介导转染人肺癌细胞SPC-A1,用MTT检测其对细胞增殖的抑制作用;Annexin VFITC-PI结合流式细胞仪检测细胞凋亡率;PI结合流式细胞仪检测细胞周期的变化;对转染细胞的表达产物进行Western blot分析。结果NS1A蛋白能抑制人肺癌细胞SPC-A1增殖且呈时间依赖性。随着作用时间延长,重组质粒转染组细胞凋亡率可达38.17%,细胞周期中G0/G1比例升高,G1期阻滞。Western blot分析显示,重组质粒转染组细胞在相对分子质量约30000处可见特异性反应条带,与NS1A蛋白产物大小相符。结论禽流感病毒NS1A蛋白能够抑制SPC-A1细胞增殖,并诱导SPC-A1细胞凋亡。

Induction of Apoptosis of Human Lung Cancer SPC-A1 Cells with NS1A Protein of Avian Influenza Virus

Objective To study the influence of NS1A protein of avian influenza virus on the proliferation of human lung cancer SPC-A1 cells.Methods Transfect human lung cancer SPC-A1 cells with recombinant plasmid pVAX1-NS1A in mediation of liposome.Determine the inhibiting effect on cell proliferation by MTT method,the apoptosis rate by Annexin V FITC kit,PI and flow cytometry,and the cell cycle by PI and flow cytometry.Identify the expressed product in transfected SPC-A1 cells by Western blot.Results NS1A protein showed a time-dependent inhibiting effect on the proliferation of SPC-A1 cells.The apoptosis rate of SPC-A1 cells transfected with recombinant plasmid pVAX1-NS1A reached 38.17%,and the percentage of cells at G0/G1 stages increased as time goes on.The cycle of transfected SPC-A1 cells was blocked at stage G1.Wesetent blot showed a specific reaction band with relative molecular mass of about 30 000,which was consistent with that of NS1A protein.Conclusion The NS1A protein of avian influenza virus inhibited the proliferation and induced the apoptosis of human lung cancer SPC-A1 cells.