目的 探讨激活转录因子2(ATF2)对肺癌AG490细胞增殖及细胞周期的影响及机制.方法 采用PCR法合成ATF2片段,采用第三代慢病毒包装体系得到ATF2过表达慢病毒及空载质粒对照病毒.将体外培养的肺癌AG490细胞随机分为对照组和观察组,分别感染空载质粒对照病毒和ATF2过表达慢病毒,感染48 h时采用qRT-PCR法检测ATF2 mRNA相对表达量,采用流式细胞仪检测细胞周期;采用CCK-8试剂盒检测感染病毒第1、2、3、4天细胞增殖指数;感染48 h时采用Western blotting法检测ATF2、p53、p21蛋白相对表达量.结果 感染病毒48 h时,对照组及观察组ATF2 mRNA相对表达量分别为0.42±0.05、1.00±0.12;ATF2 蛋白相对表达量分别为0.23±0.04, 0.76±0.13,两组比较P均<0.01.对照组及观察组感染病毒第4天细胞增殖指数比较P<0.01,其余时间点比较P均>0.05.感染病毒48 h时,对照组及观察组 G0/G1期细胞比例分别为79.7%、66.6%,S期分别为12.6%、21.5%,G2期分别为5.4%、11.4%,subG1期分别为2.3%、0.5%,两组G0/G1、S、G2细胞比例比较P均<0.05.感染病毒48 h时,对照组及观察组p53蛋白相对表达量分别为2.7±0.4、1.6±0.2,p21蛋白相对表达量分别为1.8±0.3、0.2±0.0;两组比较P均<0.01.结论 ATF2可促进肺癌AG490细胞增殖、细胞周期活化;抑制p53、p21蛋白表达可能是其作用机制.%Objective To investigate the effect of activating transcription factor 2 (ATF2) on cell proliferation and cell cycle of lung carcinoma AG490 cells.Methods Using PCR to synthesize the ATF2, employing third generation lentivirus packaging system to get the control virus and ATF2 overexpression virus.The lung cancer cell line AG490 was randomly divided into the control group and the observation group, which was infected by the control virus and ATF2 overexpression virus.Using qRT-PCR to detect the expression of ATF2 in mRNA level, and the cell cycle was detected by flow cytometry.The cell proliferation index was detected by CCK-8 kit at 1, 2, 3 and 4 days after infection.The relative expression of ATF2, p53 and p21 protein was detected by Western blotting at 48 h.Results After infecting the lung carcinoma cell for 48 h by using control and ATF2 virus, the expression level of ATF2 mRNA and ATF2 protein increased in the observation group as compared with that of the control group (all P<0.01).The proliferation index on the day 4 of the observation group was higher than that in the control group (P<0.05).The expression of P53 and P21 protein in observation group was higher than that in the control group (both P<0.01).Conclusion ATF2 can promote the cell proliferation, activation of the cell cycle in lung carcinoma cell line AG490 by inhibiting P53 and P21 protein expression.
展开▼
