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目的探讨非CO2依赖型细胞培养系统对膀胱癌T-24细胞增殖的影响。方法对膀胱癌T-24细胞采取非CO2依赖型细胞培养系统在(37℃、大气环境)培养箱(大气-B组)和(37℃、5%CO2)培养箱(CO2-B组)条件下,及CO2依赖型细胞培养系统在(37℃、大气环境)培养箱(大气-A组)和(37℃、5%CO2)培养箱(CO2-A组)条件下进行培养,检测分析两种体系培养基连续7d pH的比较结果;采用细胞增殖曲线及克隆形成实验检测细胞生长差异情况;采用流式细胞术检测分析两种系统对细胞周期及凋亡率影响的比较结果;RT-PCR、Western blot技术检测分析两种系统下的膀胱癌细胞中survivin、caspase-3基因及蛋白的表达情况。结果两种培养系统条件下,细胞培养基pH变化趋势一致;两种培养系统下T-24细胞生长增殖未见差异;RT-PCR法及Western bolt法检测显示,两种培养系统下表达survivin、caspase-3因子及蛋白未见差异。结论膀胱癌非CO2依赖型细胞培养系统对细胞增殖、基因表达等方面与膀胱癌CO2依赖型细胞培养系统无明显区别。
Objective To investigate the effects of non-CO2-dependent cell culture system on the proliferation of bladder cancer T-24 cells. Methods The bladder cancer T-24 cells were cultured in a non-CO2-dependent cell culture system (atmosphere-B) and CO2 incubator (37 ° C, 5% CO2) (CO2-dependent cell culture system) under the conditions of (37 ℃, 5% CO2) incubator (atmosphere-A group) and CO2-A group The results of the comparison of the cell growth curve and the colony formation assay were used to detect the cell growth difference. The flow cytometry was used to analyze the effects of the two systems on cell cycle and apoptosis rate. RT-PCR Western blot was used to detect the expression of survivin and caspase-3 gene and protein in the two kinds of bladder cancer cells. Results The cell culture medium showed the same trend of pH change under the two culture conditions. There was no difference in T-24 cell proliferation between the two culture systems. The expression of survivin, There was no difference in caspase-3 and protein. Conclusion The non-CO2-dependent cell culture system of bladder cancer has no significant difference from the cell-free culture system of CO2-dependent bladder cancer in terms of cell proliferation and gene expression.