DCX和SPARC基因共表达增强X射线对人脑胶质瘤细胞U-87MG凋亡的影响

Effects of DCX and SPARC co-expression on the apoptosis of human glioma cells U-87MG induced by X-rays irradiation

用激光共聚焦显微镜观察绿色荧光蛋白(GFP)的表达、流式细胞仪检测GFP表达比例确定重组腺病毒对U-87MG细胞的最佳感染剂量;RT-PCR和Western blot法检测DCX和SPARC在U-87MG细胞mRNA和蛋白水平的表达;流式细胞仪检测Ad-GFP、Ad-DCX、Ad-SPARC和Ad-DCX-SPARC对X射线诱导U-87MG细胞凋亡的影响。实验结果表明:重组腺病毒对U-87MG细胞的最佳感染剂量为50 MOI;目的基因DCX和SPARC可在mRNA和蛋白水平成功表达;Ad-GFP、Ad-DCX、Ad-SPARC和Ad-DCX-SPARC单纯基因组的凋亡率分别为(1.05±0.42)%、(1.11±0.29)%、(1.18±0.37)%和(1.12±0.40)%;8 Gy剂量辐照后,凋亡率分别为(7.54±0.47)%、(23.23±2.72)%、(16.43±1.13)%和(33.01±2.67)%。提示Ad-DCX-SPARC可以增强X射线诱导的U-87MG细胞的凋亡,从而增强放射敏感性。

The laser confocal microscope and flow cytometry （FCM） were used to confirm the best infection dose. The expression on mRNA and protein levels of DCX and SPARC were detected by RT-PCR and Western blot. FCM was used to detect the apoptosis rate. The results showed that the best infection dose of U-87MG cells was 50MOI. DCX and SPARC were successfully expressed in U-87MG cells. Without irradiation, the apoptosis rates of Ad-GFP group, Ad-DCX group, Ad-SPARC group, and Ad-DCX-SPARC group were （1.05±0.42）%, （1.11±0.29）%, （1.18±0.37）%, and （1.12±0.40）%, respectively. And after 8 Gy dose irradiation, the apoptosis rates were （7.54±0.47）%, （23.23±2.72）%, （16.43±1.13）%, and （33.01±2.67）%, respectively. The results revealed that Ad-DCX-SPARC could augment the apoptosis induced by X-rays irradiation.