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目的 :探讨低氧微环境对人脑胶质瘤U251细胞体外生长、增殖、周期分布及凋亡的影响。方法 :将U251细胞置于不同氧浓度(常氧、3%氧浓度和1%氧浓度)条件下培养,然后倒置相差显微镜下观察各组细胞的形态差异,CCK-8法和平板克隆形成实验分别检测各组细胞的增殖活性及克隆形成能力,FCM法检测各组细胞的周期分布及凋亡情况。结果 :3%低氧组U251细胞较常氧组和1%低氧组生长状态更好。3%低氧组U251细胞的增殖活性和克隆形成能力均明显优于常氧组,而常氧组又优于1%低氧组,差异均具有统计学意义(P值均<0.05)。3%低氧组U251细胞中增殖期(S+G2)细胞所占比例明显高于常氧组,常氧组又高于1%低氧组(P值均<0.05);而3%低氧组细胞的凋亡率明显低于常氧组,常氧组又低于1%低氧组(P值均<0.05)。结论 :适度的低氧微环境能促进胶质瘤U251细胞的体外生长和增殖,提高增殖期细胞的比例,抑制细胞凋亡。
Objective: To investigate the effects of hypoxic microenvironment on the growth, proliferation, period distribution and apoptosis of human glioma U251 cells in vitro. Methods: U251 cells were cultured under different oxygen concentrations (normoxia, 3% oxygen and 1% oxygen). The morphological differences of the cells were observed under inverted phase contrast microscope. The CCK-8 assay and plate clone formation assay The proliferative activity and clonogenic capacity of each group were detected respectively. The cell cycle distribution and apoptosis of each group were detected by FCM. Results: U251 cells in 3% hypoxia group grew better than those in normoxia group and 1% hypoxia group. The proliferative activity and clonogenic capacity of U251 cells in 3% hypoxia group were significantly better than those in normoxia group, while those in normoxia group were superior to 1% hypoxia group (all P <0.05). The proportion of cells in proliferating phase (S + G2) of U251 cells in 3% hypoxia group was significantly higher than that in normoxia group and that in normoxia group was higher than that in 1% hypoxia group (P <0.05) The apoptotic rate of the group of cells was significantly lower than that of normoxia group and that of normoxic group was lower than that of hypoxia group (P <0.05). Conclusion: Moderate hypoxic microenvironment can promote the growth and proliferation of glioma U251 cells in vitro, increase the proportion of cells in proliferating phase and inhibit apoptosis.