摘要
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 evaluate the effects of the CO2-independent cell culture system on the proliferation of bladder cancer cells T-24.Methods Bladder cancer cells T-24 were cultured in the CO2-independent cell culture system at 37 ℃,in a 5% carbon dioxide incubator（CO2-B group） and in an air incubator（air-B group）,and in the CO2-dependent cell culture system at 37 ℃,in a 5% carbon dioxide incubator（CO2-A group） and in an air incubator（air-A group）.The pH values of these two cell culture mediums were measured for 7 consecutive days.Differences of the cell viability were assayed with MTS and the colony forming assay.The cell cycle and apoptosis ratio of T-24 cells treated in the two cell culture systems were tested by FCM.Expressions of survivin mRNA and caspase-3 mRNA in T-24 cells cultured with the two systems were detected by RT-PCR,while expressions of survivin and caspase-3 proteins were detected by Western blot.Results The pH trends of the two cell culture mediums were the same.T-24 cell proliferation and expressions of survivin and caspase-3 mRNAs and proteins had no obvious difference.Conclusion The CO2-independent cell culture system is not obviously different from the CO2-dependent cell culture system in cell proliferation and gene expression.
出处
《山东大学学报（医学版）》
CAS
北大核心
2010年第10期60-64,共5页
Journal of Shandong University:Health Sciences
基金
桂林市科学研究与技术开发计划资助项目(20090112-4)
广西壮族自治区卫生厅自筹课题(Z2008277)
