Xiang Wei, Zhu Xianli, Zhao Hongyang
Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2005;25(4):448-50. doi: 10.1007/BF02828220.
To evaluate the effect of wild-type p53 gene on the growth and radiotherapeutic sensitivity of human glioma cells, plasmid PC53-SN3 carrying wild-type p53 gene was transfected into U251 cells. p53 gene expression in transfected cells was detected by RT-PCR, and the cell growth inhibition and apoptosis in the absence or presence of irradiation were assessed by MTT and flow cytometry. The transfection of p53 gene into U251 cells was confirmed by RT-PCR. MTT showed that p53 gene alone induced strong inhibitory effect on the growth of U251 cells (inhibition rate (IR), (79.60 +/- 5.69)%). The killing effect of irradiation alone on U251 cells was not strong (IR: (17.06 +/- 4.35)% (17.39 +/- 1.67)% (18.73 +/- 4.68)%) and increased with the irradiation doses (3, 6, 9 Gy). When combined treatment of wild-type p53 gene transfection and irradiation was used, the effect was significantly increased (IR:(80.60 +/- 5.35)%. (90.30 +/- 1.67)%, (91.30 +/- 2.01)%). The apoptosis rate of U251 cells induced by p53 gene transfection was 17.38%. The rate induced by irradiation increased (4.61%, 4.84%, 5.40%) with the irradiation doses (3, 6, 9 Gy). The apoptosis rate was also significantly increased (17.80%, 20.03%, 22.34%) after combined treatment of p53 and irradiation with different doses (3, 6, 9 Gy). It is concluded that wild-type p53 gene and irradiation could result in synergistic inhibitory effect on the growth of human glioma cells.
为评估野生型p53基因对人胶质瘤细胞生长及放射治疗敏感性的影响,将携带野生型p53基因的质粒PC53-SN3转染至U251细胞。采用RT-PCR检测转染细胞中p53基因的表达,通过MTT法和流式细胞术评估在有无照射情况下细胞的生长抑制及凋亡情况。RT-PCR证实p53基因已转染至U251细胞。MTT结果显示,单独p53基因对U251细胞生长具有较强抑制作用(抑制率(IR)为(79.60±5.69)%)。单独照射对U251细胞的杀伤作用不强(IR分别为(17.06±4.35)%、(17.39±1.67)%、(18.73±4.68)%),且随照射剂量(3、6、9 Gy)增加而增强。当采用野生型p53基因转染与照射联合治疗时,效果显著增强(IR分别为(80.60±5.35)%、(90.30±1.67)%、(91.30±2.01)%)。p53基因转染诱导U251细胞的凋亡率为17.38%。照射诱导的凋亡率随照射剂量(3、6、9 Gy)增加而升高(分别为4.61%、4.84%、5.40%)。p53与不同剂量(3、6、9 Gy)照射联合治疗后,凋亡率也显著升高(分别为17.80%、20.03%、22.34%)。结论:野生型p53基因与照射对人胶质瘤细胞生长可产生协同抑制作用。