【摘要】 采用RT?PCR(逆转录聚合酶链反应)半定量方法检测p16基因转录水平的变化,采用免疫细胞化学方法检测蛋白表达,观察X射线照射对离体培养的EL?4细胞p16基因转录及蛋白表达的影响,证实p16基因转录表达在辐射诱导EL?4细胞G1期阻滞中的重要作用。结果表明,2.0GyX射线照射后2—48h,EL?4细胞p16mRNA水平增高,照射后72h恢复正常水平。P16蛋白表达于照射后2h开始增高,照射后12hP16蛋白表达非常显著高于对照组(p<0.01)。剂量?效应实验表明,0.5—6.0GyX射线照射后12h,?4细胞p16mRNAEL水平明显提高。2.0、4.0、6.0Gy组P16蛋白表达均非常显著增加(分别p<0.01)。研究证实,电离辐射能够诱导EL?4细胞p16基因转录及蛋白表达增高,其增幅呈现明显的剂量依赖性。提示辐射诱导p16基因转录表达增加在辐射诱导EL?4细胞G1期阻滞中发挥重要作用。更多还原

【Abstract】 To investigate the effect of ionizing radiation on p16 gene transcription and protein expression in EL-4 cells in vitro, RT-PCR semi-quantity method was applied to determine the p16mRNA level and immunocytochemistry (ICC) was employed to measure the p16 protein expression. The time-course experiments showed that the p16 mRNA level increased at 2—48h in EL-4 cells, and the p16 protein expression increased significantly in EL-4 cells (p<0.01) at 12h after 2.0Gy X-irradiation. The dose-effect experiments showed that the p16mRNA level increased with the dose, and the p16 protein expression increased significantly (p<0.01) with the dose in 2.0 - 6.0Gy range. The results suggested that the increase of p16 gene transcription and protein expression in EL-4 cells might play role in G1 arrest induced by ionizing radiation in vitro.更多还原