细胞微缺氧模型的建立及其对胃癌细胞生物学行为的影响

Establishment of Cellular Hypoxia Model and Its Effects on Biological Behavior of Gastric Cancer Cell Line SGC-7901

背景与目的:缺氧是实体瘤微环境的基本特征之一,体外制造微缺氧的细胞培养环境,对研究缺氧对肿瘤细胞的影响有重要意义。目前建立体外缺氧细胞培养模型的方法较为烦琐,尚缺少微缺氧对胃癌细胞影响较为完整的研究。本实验运用GasPaK产气袋法体外创建胃癌细胞微缺氧培养模型,并观察该状态下细胞生物学行为的变化。方法:运用产气袋法(GasPaK)制造微缺氧条件,血气分析监测RPMI-1640培养液中PO2、PCO2和pH值变化。流式细胞仪分析微缺氧对SGC-7901细胞周期分布的影响,光镜及透射电镜观察微缺氧条件下SGC-7901细胞形态的改变,台盼蓝染色计数微缺氧对活细胞数的影响,MTT法检测微缺氧对SGC-7901细胞粘附能力的影响,游走实验检测微缺氧对SGC-7901细胞游走运动能力的影响。结果:GasPaK产气袋法在0.5～48h内PO2、PCO2和pH稳定,并能达到微缺氧细胞培养的条件。微缺氧处理SGC-7901细胞16h后未见细胞周期的变化。微缺氧导致部分SGC-7901细胞形态发生改变。微缺氧处理16h后活细胞计数无明显下降,微缺氧组SGC-7901细胞粘附能力增强,游走穿膜细胞数(196±9)是对照组(114±7)的1.71倍(P<0.05)。结论:通过GasPaK产气袋法制造的微缺氧肿瘤细胞培养环境,达到了条件稳定、重复性好的特点。微缺氧对SGC-7901细胞有影响,但这种影响相对较小,SGC-7901细胞对微缺氧有一定的耐受性。

BACKGROUND ＆ OBJECTIVE： Hypoxia is an elementary characteristic of tumor microenvironment. Establishing hypoxia microenvironment in vitro will be of help in studying the effects of hypoxia on tumor cells. Previous methods of establishing hypoxic cell culture model are fussy, and researches on the effects of hypoxia on gastric cancer cells are rare. This study was to establish hypoxia microenvironment with GasPaK method in vitro, and observe the effects of hypoxia on biological features of gastric cancer cell line SGC-7901. METHODS： The hypoxia microenvironment was established by GasPaK method; the changes of PO2, PCO2, and pH in RPMI-1640 were monitored by blood gas analysis. The effect of hypoxia on cell cycle of SGC-7901 cells was analyzed by flow cytometry （FCM）. The morphology of SGC-7901 cells was observed under light microscope and transmission electron microscope. Live cells were counted with trypan blue staining, The adhesiveness of SGC-7901 cells was detected by MTT assay; migration ability of SGC-7901 cells was assessed by movement experiment, RESULTS： GasPaK formed hypoxia microenvironment in 0.5-48 h with stable PO2, PCO2, and pH. When cultured in hypoxia for 16 h, no significant change in cell cycle of SGC-7901 cells was observed; morphology of some SGC-7901 ceils was changed; the number of live cells didn＇t significantly decreased; the adhesiveness and migration ability of SGC-7901 cells were significantly enhanced. CONCLUSIONS： GasPaK method can stably establish hypoxia microenvironment with good repetition. Hypoxia has slight effect on SGC-7901 cells, and may induce morphologic change of SGC-7901 cells.