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目的 探讨X 刀治疗脑胶质瘤的最佳剂量—效应关系和最佳时间—效应关系。方法 以BT3 2 5细胞系为实验对象 ,设置了阴性对照组、5Gy、10Gy、15Gy、2 0Gy和 3 0Gy组 ,应用Varian直线加速器行单次大剂量照射 ,分别于 2h、6h、12h、4 8h、72h和 96h采用流式细胞仪观察细胞凋亡和细胞周期的改变 ;采用免疫组化法观察p5 3、BcL 2和PCNA基因的表达情况 ;采用光镜和电镜观察细胞凋亡的形态学改变。结果 X线诱导BT3 2 5细胞系凋亡的最佳剂量为 2 0Gy ,凋亡率为 66 95 %。BT3 2 5细胞受一定剂量X线照射后凋亡细胞逐渐增加 ,12～ 2 4h左右到平台 ,以后即使再增加剂量 ,细胞凋亡也不再增加。在 2 0Gy 4 8h时 ,p5 3基因表达最高 ,BcL 2基因表达明显减弱 ,PCNA 12h内阳性率较高 ,随着时间延长 ,其表达率明显下降 ,尤以 4 8h为著。且细胞周期发生G1 阻滞 ,达 85 5 6% ,此时经FCM检测细胞凋亡发生率也最高。结论 X线诱导细胞系凋亡存在最佳剂量—效应关系和最佳时间—效应关系 ,且与p5 3、BcL 2和PCNA基因的表达有一定的关系
Objective To explore the best dose-response relationship and optimal time-effect relationship of X knife in treating glioma. Methods The BT3 2 5 cell line was used as the experimental group, and the negative control group, 5Gy, 10Gy, 15Gy, 20Gy and 30Gy groups were set up and irradiated with single high dose of Varian linear accelerator at 2h, 6h, 12h, 4 8h, 72h and 96h by flow cytometry apoptosis and cell cycle changes; using immunohistochemistry p5 3, BcL 2 and PCNA gene expression; using light microscopy and electron microscopy apoptosis morphology change. Results The optimal dose of X-ray irradiation for inducing apoptosis in BT3 2 5 cells was 20 Gy and the apoptosis rate was 66 95%. BT3 2 5 cells under certain dose of X-ray irradiation apoptotic cells increased gradually, about 12 ~ 2 4h to the platform, even after further increase the dose, apoptosis is no longer increased. The expression of p5 3 gene was highest at 2 0 Gy 4 h and the expression of BcL 2 gene was significantly weakened. The positive rate of PCNA within 12 h was higher than that of PCNA. The expression of p5 3 was significantly decreased with the prolongation of time, especially 48 h. G1 arrest occurred in the cell cycle, reaching 85.56%. At this time, the apoptosis rate by FCM was the highest. Conclusion There is the best dose-effect relationship and the best time-effect relationship between X-ray-induced cell apoptosis and the expression of p53, Bcl-2 and PCNA genes