X射线对人肺癌细胞Pokemon表达影响

目的 研究不同剂量X射线照射及照射后不同时间点对体外培养的人肺癌A₅₄₉细胞系Pokemon基因和蛋白表达的影响。方法 用X射线2、4、6、8 Gy吸收剂量照射体外培养的A₅₄₉细胞系,分别采用荧光实时定量PCR技术和蛋白印记法检测A₅₄₉细胞照射后2、4、8、12、24、48 h的Pokemon mRNA和蛋白表达水平,以未照射组为对照。采用二苯基四氮唑溴盐法分别检测2 Gy照射1、2、3、4、5 d后A₅₄₉细胞增殖能力变化,以未照射组为对照。结果 Pokemon mRNA的表达在2、4、6、8 Gy照射后早期呈降低趋势,但晚期呈升高趋势,大部分时间点差异有统计学意义(t=3.40~154.76,P=0.000~0.041);Pokemon 蛋白的表达较对照组均升高,且各剂量组均在照射后8 h达峰值,在大部分时间点实验组与对照组差异有统计学意义(t=4.18~89.64,P=0.000~0.039)。照射组细胞接种后第3~5天细胞增殖能力显著低于对照组(t=2.34~18.19,P=0.000~0.040)。结论 一定剂量X射线在特定条件下可诱导人肺癌A₅₄₉细胞Pokemon mRNA和蛋白表达升高,可能与A₅₄₉细胞辐射抗性有关。

Effects of X-ray irradiation on the expression of Pokemon gene in human lung adenocarcinoma cell line

Objective To study the dose and time effects of X-ray radiation on the expression of Pokemon gene and protein in human lung adenocarcinoma cell line A549.Methods A549 cells was exposed to different doses of X-ray (2,4,6 and 8 Gy),and the expression of Pokemon mRNA and protein of the cells was detected by using Quantitative real-time PCR and western-blotting at 2,4,8,12,24,and 48 h after irradiation. 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazolium bromide was used to detect the proliferation of A549 cells at 1,2,3,4,and 5 d after 2 Gy X-ray irradiation.The mock treated A549 cells were used as the control.Results The expression of Pokemon mRNA trended to decrease after irradiated with 4,6 and 8 Gy in the earlier period and increased in the later period with statistical difference at the most time points (t =3.40 -154.76,P =0.000 -0.041 ).The expression of Pokemon protein trended to increase and reached the peak at 8 h after irradiated of 2,4,6 and 8 Gy with statistical difference at the most time points ( t =4.18 - 89.64,P =0.000 - 0.039).Compared with the control,the proliferation of A549 cells was significantly inhibited during 3 to 5 d after irradiation of 2 Gy ( t =2.34 - 18.19,P =0.000 -0.040).Conclusions X-ray irradiation may increase the expression of Pokemon mRNA and protein in A549 cells,which might be correlated with radiation-resistance of A549 cells.