缺氧增强胶质瘤干细胞“干性”的初步研究

Hypoxia enhances stemness of cancer stem cells in glioblastoma

目的研究在体外环境下,缺氧与肿瘤干细胞(CSCs)"干性"的关系。方法选择U87细胞和U251细胞以及原代胶质瘤细胞并进行缺氧处理。用透射电子显微镜法来检测这些细胞的超微结构;用MTT法来检测细胞生长;用流式细胞仪来检测细胞周期以及CD133的表达;用transwell法来检测细胞迁移能力;用集落形成分析来分析细胞的集落形成能力;并用实时定量PCR来检测干细胞以及其分化标志物的m RNA表达。结果缺氧使原代胶质瘤细胞维持在一个未分化的状态,减慢处于相对静止阶段的胶质瘤细胞的生长、增加它们的集落形成能力和胶质瘤细胞的转移,而且能提高干细胞标志物的表达水平。肿瘤干细胞的分化标志物在进行缺氧处理后降低。结论缺氧能够使已分化的胶质瘤细胞"去分化"而使其获得"干性"。

Objective To investigate the relationship between hypoxia and stemness of cancer stem cells（ CSCs）. Methods U87 cells,U251 cells and primary glioma cells were experienced hypoxia. Detected the ultrastructure of these cancer cells with transmission electron microscopy; detect the cell growth with MTT assay; cell cycle and CD133 expression were detected by flow cytometry; and the cell migration ablity were detected through transwell chamber assay; the colony-forming efficiency were deteced by colony-forming assay; and realtime quantitative PCR and Western blot were carried out to detect the m RNA and protein expression of markers of stem cells and their differentiation respectively. Results Hypoxia maintained the undifferentiated state of primary glioma cells,slowed down the growth of glioma cells which were in a relatively quiescent stage,increased the colony forming efficiency and migration of glioma cells,and increased the expression of markers of stem cells,but the expression of markers for stem cell differentiation was reduced after hypoxia treatment. Conclusion Hypoxia may induce the ＂dedifferentiation＂of differentiated glioma cells which then acquire the stemness.