慢病毒介导FGFR1沉默对肺癌细胞增殖和凋亡的影响

Effect of lentivirus mediated FGFR1 silence on proliferation and apoptosis of lung cancer cells

目的探讨慢病毒介导成纤维生长因子受体1(FGFR1)沉默对肺癌细胞增殖和凋亡的影响。方法构建慢病毒表达载体LV-shFGFR1,对其进行包装和病毒颗粒制备。将A549肺癌细胞分为干扰组、空载体组及对照组。采用实时荧光定量聚合酶链反应和Western blotting检测各组细胞FGFR1 mRNA和蛋白相对表达量,MTT比色法检测细胞增殖情况,流式细胞术和Hoechest染色检测细胞凋亡情况。结果①3株肺癌细胞中,A549细胞的FGFR1 mRNA相对表达量高于H3255和A-427细胞(P<0.05)。②3组A549细胞中FGFR1 mRNA和蛋白的相对表达量比较,差异有统计学意义(P<0.05);空载体组与对照组比较,差异无统计学意义(P>0.05);干扰组低于对照组(P<0.05)。③干扰组、空载体组、对照组0、12、24、48和72 h的A549细胞增殖情况比较,不同时间点、各组间、随时间变化趋势均有差异(P<0.05)。④3组A549细胞凋亡率比较,差异有统计学意义(P<0.05);空载体组与对照组比较,差异无统计学意义(P>0.05);干扰组高于对照组(P<0.05)。结论慢病毒介导FGFR1沉默抑制A549肺癌细胞增殖,同时促进肺癌细胞凋亡,提示FGFR1可能参与肺癌细胞的发生、发展过程。

Objective To explore the effect of lentivirus mediated FGFR1 silence on proliferation and apoptosis of lung cancer cells.Methods Lentiviral vector LV-shFGFR1 was established,packaged,and virus particle was prepared.The experiment was divided into three groups,the interference group,empty load group,and control group.FGFR1 protein and mRNA expression levels in the three groups cells were detected by using western blot and real time fluorescence quantitative technique.Cell proliferation in each group was tested by using methyl thiazolyl tetrazolium and cell apoptosis was detected by using flow cytometry and Hoechest staining.Results①The expression of FGFR1 mRNA in A549 was higher than that of H3255 and A-427(P<0.05).②The relative expression of FGFR1 mRNA and protein in A549,H3255 and A-427 lung cancer cells was significantly different(P<0.05);There was no significant difference in FGFR1 mRNA and protein expression between empty vector group and control group(P>0.05).The expression of FGFR1 mRNA and protein in the interference group was lower than that in the control group(P<0.05).③The proliferation of A549 cells in the interference group,the empty vector group and the control group at 0 h,12 h,24 h,48 h and 72 h were significantly different(P<0.05).④The apoptosis rates of A549 cells in three groups were statistically different(P<0.05);there was no significant difference in apoptosis rate between empty load group and control group(P>0.05);the apoptosis rate of A549 cells in the interference group was higher than that in the control group(P<0.05).Conclusion Lentivirus mediated FGFR1 silence can inhibit the proliferation of lung cancer cell A549 and promote the apoptosis of lung cancer cells simultaneously.It is suggested that FGFR1 may be involved in the process of lung cancer cell development.