Effect of XAF1 gene on lung adenocarcinoma A549 cell apoptosis induced by cisplatin

OBJECTIVE:Based on the apoptosis process of A549cells induced by cisplatin,we researched the expression and functions of the XAF1gene.METHODS:The proliferation inhibition rate after A549cells treated with the cisplatin of 4μg/mL was detected by MTT.The A549cell apoptosis was detected by AnnexinⅤ/7-AAD.The XAF1mRNA and protein expression was monitored by the RT-PCR and western blotting.The Caspase-3activity in apoptotic A549cells was detected by the Caspase-3assay.RESULTS:The cell proliferation inhibition rate increased with the cisplatin treating duration.The cell proliferation inhibition rates of the experiment group were 2h(3.40±0.65)%,12h(8.31±0.73)%,and 24h(14.72±0.24)%respectively.Compared with the cell proliferation inhibition rate(1.03±0.28)%of the control group,Pvalues were 0.004,0.000,and 0.000respectively.The cell apoptosis rate increased with the cisplatin treating duration.The apoptosis rates were 2h(4.35±0.95)%,12h(9.69±2.60)%,and 24h(22.35±1.24)% respectively.Compared with the cell apoptosis rate(1.39±0.21)% of the control group,Pvalues were 0.006,0.005,and 0.000respectively.The mRNA expressions of the experiment group were 2h(0.199±0.029),12h(0.654±0.093),and 24h(1.216±0.101)respectively,while the mRNA expression of the control group was(0.091±0.020).The XAF1mRNA expression increased with the cell apoptosis rate.Compared with the mRNA expression of the control group,Pvalues were0.006,0.001,and 0.000respectively.The XAF1protein expressions of the experiment group were 2h(0.322±0.041),12h(0.508±0.014),and 24h(0.901±0.014)respectively,while the XAF1protein expression of the control group was(0.124±0.007).The protein expression of the experiment group was increased with the cell apoptosis rate.Compared with the XAF1protein expression of the control group,Pvalues were 0.001,0.000,and 0.000respectively.The Caspase-3activity increased with the cisplatin treating duration.The Caspase-3activity values of the experiment group were 2h(34.745±3.781),12h(69.524±3.096),and 24h(94.787±5.429)respectively.The Caspase-3activity value of the control group were(21.914±1.962).CONCLUSIONS:A549cell apoptosis induced by cisplatin may increase the XAF1gene expression.The expression level increases according to the improvement of A549cell apoptosis level,which indicates that the XAF1gene takes effects in the apoptosis process of A549cells induced by cisplatin.