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目的:探讨模拟微重力(Modeled microgravity,MMG)对恶性胶质瘤细胞U87形态、生长增殖、基质金属蛋白酶-2、-9(Matrix metalloproteinase 2,MMP-2、Matrix metalloproteinase 9,MMP-9)及神经胶质酸性蛋白(Glial fibrillary acidic protein,GFAP)表达的影响。方法:常规培养U87细胞,传代培养3代后分为两组,正常重力组(Normal gravity,NG组)及模拟微重力干预组(Modeled microgravity,MMG组),相应条件下培养24 h,48 h和72 h。倒置显微镜观察U87细胞形态改变,细胞计数法及四甲基偶氮唑盐微量酶反应比色法(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT法)检测模拟微重力干预和干预后U87细胞生长增殖情况;Western Blot检测不同培养时间后胶质瘤细胞U87 MMP-2、MMP-9及GFAP蛋白的表达情况。结果:模拟微重力条件下培养72 h后,U87细胞轮廓不规则,边缘圆钝,触角变少;模拟微重力条件下,U87细胞生长速度明显减慢,并且经模拟微重力干预48 h和72 h后的胶质瘤细胞经传代再培养,其增殖率也明显低于正常重力组;同时,U87细胞MMP-2及MMP-9蛋白表达水平与模拟微重力处理时间呈时间依赖性下降,而GFAP蛋白表达水平则呈时间依赖性升高。结论:模拟微重力影响U87细胞的形态、生长及表型。
Objective: To investigate the effects of simulated microgravity (MMG) on the morphology, growth and proliferation, matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) Glial fibrillary acidic protein (GFAP) expression. Methods: U87 cells were cultured routinely and subcultured for 3 passages, then divided into two groups: normal gravity group (NG group) and Modeled microgravity group (MMG group), cultured under the corresponding conditions for 24 h and 48 h And 72 h. The morphological changes of U87 cells were observed by inverted microscope, and the cell counting method and MTT method were used to detect the content of U87 cells. Detect the growth and proliferation of U87 cells after simulated microgravity intervention and intervention; Detect the expression of U87 MMP-2, MMP-9 and GFAP protein in glioma cells with different culture time by Western Blot. RESULTS: U87 cells were irregular in outline, with blunt edges and few antennae after 72 h of culture under simulated microgravity. Under simulated microgravity, the growth rate of U87 cells was significantly slowed down. After simulated microgravity intervention, 48 h and 72 h after glioma cells were subcultured, the proliferation rate was significantly lower than normal gravity group; the same time, U87 cells MMP-2 and MMP-9 protein expression levels and simulated microgravity treatment time-dependent decline, and GFAP protein expression increased in a time-dependent manner. Conclusion: The microgravity influences the morphology, growth and phenotype of U87 cells.