摘要目的 研究NADPH氧化酶(NOX)对U251胶质瘤细胞存活、增殖和凋亡的影响.方法 RT-PCR检测U251胶质瘤细胞系NOX基因的表达.分别使用5、15、25 μmol/LNOX抑制剂DPI及10 mmol/L抗氧化剂Tiron处理U251细胞,24h后alamarBlue法检测U251细胞的增殖,流式细胞术检测细胞内活性氧的产生和U251细胞的凋亡情况,并与正常对照组(不做任何处理的1的U251细胞进行比较.结果 U251细胞系中明显表达NOX4 mRNA.各浓度DPI及10 mmol/Ltiron均能抑制U251胶质瘤细胞的生长,诱导U251细胞凋亡.与正常对照组比较,各浓度DPI处理组的U251细胞内的活性氧簇(ROS)均明显减少,差异有统计学意义(P<0.05); 结论 NOX4可能是胶质瘤细胞内ROS生产的主要来源.NOX4可能通过增加细胞内ROS水平并作用于其下游调节分子,对胶质瘤细胞的生长、存活和凋亡起着重要的调节作用.

abstractsObjective To investigate the effects of NADPH oxidase (NOX) on the survival,proliferation and apoptosis of U251 glioma cells. Methods RT-PCR was employed to examine the expressions of NOX genes in the U251 glioma cells. Five, 15 and 25 μmol/L diphenyleneiodonium (DPI,the NOX inhibitor) and 10 mmol/L antioxidant Tiron were added into the cells, respectively, and 24 h after that, the proliferation of U251 glioma cells was tested by alamarBlue assay, and the production of intraceilular reactive oxygen species (ROS) and the apoptosis of U251 glioma cells were examined by flow cytometry. These results were compared with those in the normal control group. Results High mRNA expression level of NOX4 gene was found in the U251 glioma cells. DPI at each concentration can inhibit the growth of U251 glinma cells and induce the apoptosis of U251 glioma cells. Compared with the normal control group, the treatment groups showed significantly decreased ROS in the U251 glioma cells (P<0.05). Conclusion NOX4 gene may be a major source that generates the intracellular ROS in the glioma cells. NOX4 gene regulates the proliferation, survival and apoptosis of glioma cells through increasing the level of ROS and then acting on its downstream regulatory molecules.