GRHL2沉默对肺癌A549细胞增殖及凋亡的影响

目的探讨GRHL2沉默对肺癌A549细胞的增殖、凋亡的影响。方法利用RNA干扰技术,构建GRHL2沉默慢病毒表达载体,将肺癌A549细胞分为3个组,实验组:GRHL2沉默慢病毒感染的A549细胞系;阴性对照组:空载慢病毒感染的A549细胞系;空白对照组:不给予任何处理的A549细胞系。采用倒置荧光显微镜观察细胞绿色荧光蛋白的强度,通过实时定量聚合酶链式反应和蛋白质印迹法检测肺癌A549细胞中GRHL2 mRNA和蛋白质表达水平。采用细胞计数试剂盒检测GRHL2沉默对A549细胞增殖的影响。采用Annexin V-APC/PI细胞凋亡检测试剂盒及流式细胞术分析GRHL2沉默对A549细胞凋亡的影响。结果实验组中GRHL2 mRNA及蛋白质表达水平均较空白对照组及阴性对照组显著降低(F=48.13、42.57,P值均<0.05),A549细胞的增殖能力明显下降(F=65.64,P<0.05),凋亡显著增强(F=56.76,P<0.05)。结论靶向沉默GRHL2能显著抑制肺癌A549细胞的增殖并促进其凋亡,GRHL2有望成为临床肺癌靶向治疗的新靶点。

Effect of silencing of GRHL2 on proliferation and apoptosis of lung cancer A549 cells

Objective To investigate the effect of GRHL2 silencing on proliferation and apoptosis of lung cancer A549 cells.Methods Using RNA interference technology,the expression vector of GRHL2 silenced lentivirus was constructed to infect lung cancer A549 cells,the experiment was divided into the experimental group(GRHL2 silences the A549 cell line infected by lentivirus),the negative control group(the A549 cell line infected by no-load lentivirus),the blank control group(the A549 cell line without any treatment).The intensity of green fluorescent protein was observed by inverted fluorescence microscope,and the expression levels of GRHL2 mRNA and protein in lung cancer A549 cells were detected by quantitative real-time polymerase chain reaction and Western blot.Cell counting kit-8 was used to detect the effect of GRHL2 silencing on A549 cell proliferation.Annexin V-APC/PI reagents and flow analysis were used to analyze the effect of GRHL2 silencing on the apoptosis of A549 cells.Results The expression levels of GRHL2 mRNA and protein in the experimental group were significantly decreased than the negative control groupand blank control group(F=48.13,42.57,both P<0.05),the proliferation ability of A549 cells was significantly decreased(F=65.64,P<0.05),and apoptosis was significantly increased(F=56.76,P<0.05).Conclusions Targeted silencing of GRHL2 can significantly inhibit the proliferation and promote apoptosis of lung cancer A549 cells,and GRHL2 is expected to become a new target for clinical lung cancer targeted therapy.