放射控导的wt-p53蛋白正反馈基因环路对肺腺癌 - 第三军医大学学报.DOC

放射控导的wt-p53蛋白正反馈基因环路对肺腺癌细胞的体外抗癌生物学效应研究 毛丽伟，廖国清1，王红梅1，陈正堂2 （1.中国人民解放军309医院肿瘤科；2.第三军医大学新桥医院全军肿瘤研究所） ［摘要］ 目的 研究体外对肺腺癌细胞放射敏感性的影响直线加速器8 MV X线400mu/min，4Gy，源皮距100cm的照射条件pE6R4-p53-EGFP/H1299转染细胞H1299(p53-/-)、pE6-p53-EGFP/H1299pR4-p53-EGFP/H1299转染细胞分析该环路对细胞周期和细胞凋亡率的影响；绘制细胞放射剂量－存活曲线，计算D0值及增敏比。4Gy 8MV-X线照射治疗组和对照组细胞后，治疗组在照射后发生明显G0/G1期阻滞(75.±1.42)%；照射后各组细胞凋亡率均高于照射前组，照射后治疗组细胞凋亡率(23.±0.21％)分别是对照组细胞的5.倍(4.±0.12)％15.67±0.32)％和2.57倍(9.23±0.15)％。分析计算pE6R4-p53-EGFP/ H1299、pE6-p53-EGFP/H1299和H1299(p53-/-)三组细胞D值分别0.91Gy、1.073 Gy和1.413 Gy，根据D0值计算SER分别为2.63和1.34放射可wt-p53正反馈基因基因环路上调wtp53蛋白表达肺腺癌细胞敏感性。A Analysis of anti-tumor biological effects of irradiation-induced wt-p53 positive circuit MAO Li-wei1，LIAO Guo-qing1，WANG Hong-mei1，CHEN Zheng-tang2 (1.Department of Oncology, the PLA 309 Hospital, Being 100091,China. 2.Cancer Research Center; Xinqiao Hospita,Third Military Medical University ; Chongqing 400037 ,China) [Abstract] Objective To study the effects of irradiation-induced wt-p53 positive feedback circuit on cell life cycle, apoptosis rate and irradiation sensitivity of adenocarcinoma of lung in vitro. Methods The therapeutic group pE6R4-p53-EGFP/H1299 cells and the other 3 control groups H1299(p53-/-)、pE6-p53-EGFP/H1299、pR4-p53-EGFP/H1299D0 and sensitive enhancement ratio (SER) index were calculated from the irradiation dose-survival curve. Results At 12h post 4Gy 8MV-X-ray irradiation, FCM analysis showed that most cells in therapeutic group arrested at G0/G1 stage (75.13±1.42)%, compared with 3 control groups (38.47±0.87)%, (62.57±0.76)%,(51.23±2.41)%, respectively. After irradiation, the cell apoptosis rate in each group was higher than that in cells without irradiation. The apoptosis rate in therapeutic group was (23.73±0.21)％, which was 5.69 folds (4.17±0.12)％, 1.51 folds(15.67±0.32)％ and 2.57 folds(9.23±0.15)％ of that in three control groups, respectively. The D0 values were 0.91Gy, 1.073 Gy and 1.413 Gy in pE6R4-p53-EGFP/H1299, pE6-p53-EGFP/H1299 and H1299 cells, respecti