导入野生型p53基因对人胶质瘤细胞生长及放疗敏感性的影响

Effect of wild-type p53 gene transfection on the growth and radiotherapeutic sensitivity of human glioma cells

目的 研究野生型p5 3基因对人胶质瘤细胞生长及放疗敏感性的影响。方法 将野生型 p5 3基因导入U 2 5 1细胞 ,通过逆转录 聚合酶链反应 (RT PCR)检测 p5 3基因的表达。U2 5 1细胞分为 4组 :对照组、转染组、放疗组、转染联合放疗组。用四甲基偶氮唑盐微量酶反应比色法(MTT法 )和流式细胞仪检测p5 3基因或 /和放疗 (3、6、9Gy)对U 2 5 1细胞生长抑制及凋亡的影响。结果 通过RT PCR证实了 p5 3基因在U 2 5 1细胞中的表达。MTT检测发现 p5 3基因对U2 5 1细胞的生长抑制率为 (79.60± 5 .69) %。当放射剂量为 3、6、9Gy时 ,U2 5 1细胞的生长抑制率分别为(17.0 6± 4.3 5 ) % ,(17.3 9± 1.67) % ,(18.73± 4.68) %。当 p5 3基因与放疗 (3、6、9Gy)联合作用时 ,抑制率分别为 (80 .60± 5 .3 5 ) % ,(90 .3 0± 1.67) % ,(91.3 0± 2 .0 1) %。p5 3基因转染所产生的U 2 5 1细胞凋亡率为 17.3 8%。放疗 (3、6、9Gy)引起的细胞凋亡率分别为 4.61% ,4.84% ,5 .40 %。当p5 3基因与放疗 (3、6、9Gy)联合作用时 ,凋亡率分别为17.80 % ,2 0 .0 3 % ,2 2 .3 4%。结论 野生型p5

Objective To evaluate the effects of wild-type p53 gene on the growth and radiotherapeutic sensitivity of human glioma cells.Methods Wild-type p53 gene was transfected into U251 cells,and p53 gene expression was detected by RT-PCR.U251 cell was divided into 4 groups:control group,transfection group,radiation group and combined treatment group.The cell growth inhibition and apoptosis in either the absence or the presence of radiation (3,6,9 Gy) were assessed by MTT and flow cytometry.Results The transfection of p53 gene into U251 cells was confirmed by RT-PCR.MTT showed that p53 gene by itself induced strong inhibition effect on the growth of U251 cells [inhibition rate,IR (79.60± 5.69)%]. The killing effect of radiation by itself on U251 cells was not strong [IR (17.06± 4.35)%, (17.39± 1.67)%, (18.73± 4.68)%] and increased with the increase of radiation doses (3,6,9 Gy).When combined treatment of wild-type p53 gene transfection and radiation was used,that was significantly increased [IR (80.60± 5.35)%, (90.30± 1.67)%, (91.30± 2.01)%]. The apoptotic rate of U251 cells induced by p53 gene transfection was 17.38%. That induced by radiation was increased (4.61%, 4.84%, 5.40%) with the increase of radiation doses (3,6,9 Gy).The apoptotic rate was also significantly increased (17.80%, 20.03%, 22.34%) after combined treatment of p53 and radiation with different doses (3,6,9 Gy).Conclusion Wild-type p53 gene and radiation could result in synergistic inhibition effect on the growth of human glioma cells.