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目的探讨MG-132诱导人脑胶质瘤细胞SHG-44损伤中钙超载的作用。方法通过传代方法培养人脑胶质瘤细胞。选取MG-132处理24小时,采用处理前(对照组)和不同浓度(5、10、15、50μmol/L)处理后的胶质瘤细胞,采用MTT比色法检测细胞活力,流式细胞仪检测细胞凋亡情况,激光共聚焦检测细胞内钙离子荧光强度。结果MTT比色法检测发现,MG-132处理24h后,SHG-44细胞活力明显下降43%(P<0.05),且随着MG-132浓度的加大,继续呈现显著下降的趋势(P<0.05)。流式细胞仪检测结果发现,与对照组比较,MG-132作用24h后,各组细胞凋亡率显著升高(P<0.05),5、10、15μmol/L组细胞调往情况相似(P>0.05),但50μmol/L细胞凋亡率升高较为明显(P<0.05)。激光共聚焦测钙离子荧光强度结果显示,与空白对照组相比钙离子荧光强度明显升高(P<0.05),5、10、15μmol/L组钙离子荧光强度相似(P>0.05),但50μmol/L细胞钙离子荧光强度明显升高(P<0.05)。结论MG-132处理可以通过激发细胞内钙超载诱导胶质瘤细胞凋亡。
Objective To investigate the effect of MG-132 on calcium overload induced by SHG-44 in human glioma cells. Methods Human glioma cells were cultured by passage method. The cells were treated with MG-132 for 24 hours and treated with different concentrations (5, 10, 15 and 50 μmol / L) before treatment. Cell viability was measured by MTT colorimetric assay. Flow cytometry Cell apoptosis was detected by laser confocal laser scanning microscope. Results MTT assay showed that the viability of SHG-44 cells was significantly decreased by 43% (P <0.05) after treatment with MG-132 for 24 hours, and the MG-132 concentration continued to show a significant decrease (P < 0.05). Flow cytometry results showed that compared with the control group, MG-132 cells apoptosis rate was significantly increased after 24h (P <0.05), 5,10,15μmol / L cells were transferred to the similar situation (P > 0.05), but the apoptosis rate in 50μmol / L group was more obvious (P <0.05). The results of laser scanning confocal microscope showed that the fluorescence intensity of calcium was significantly higher than that of blank control group (P <0.05), while the fluorescence intensity of calcium ion of 5,10,15μmol / L group was similar (P> 0.05) The calcium fluorescence intensity of 50μmol / L cells was significantly increased (P <0.05). Conclusion MG-132 treatment can induce apoptosis of glioma cells by stimulating intracellular calcium overload.