摘要目的 探讨洛伐他汀(Lov)诱导子宫内膜癌细胞株HEC-1-A G1期同步化的方法及HEC-1-A细胞解除G1期同步化后的细胞周期进程.方法 Kit-8细胞计数(CCK-8)法测定HEC-1-A细胞的倍增时间,采用10、20、30、40 μmol/L的Lov作用1×细胞倍增时间,采用流式细胞术检测G1期细胞的比例,获得G1期同步化的最佳Lov浓度;采用最佳浓度作用HEC-1-A细胞,于作用后0.5×倍增时间~2×倍增时间内,每隔4h检测同步化程度,获得最佳浓度Lov作用下的最佳同步化时间;解除G1期同步化,每隔4h检测一次,研究HEC-1-A细胞解除G1期同步化后的细胞周期进程.结果 CCK-8法检测显示HEC-1-A细胞的倍增时间约为24 h;HEC-1-A细胞的最佳G1期同步化条件为:40 μmol/L Lov作用28 h,可获取G1期细胞(87.87±0.70)%;HEC-1-A细胞于解除G1期同步化后20h,获得最大量的S期细胞,为(33.58±0.62)%;同时G1期细胞比例达到最低,为( 58.42±0.54)%.结论 40 μmol/L Lov作用28 h,可获得最大量G1期细胞;解除G1期同步化后20 h,S期细胞比例最高,G1期细胞比例达到最低,达到满意的G1期同步化效果.
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abstractsObjective To investigate the effects of lovastatin on inducing G1 phase synchromzation in HEC-1-A cells and examine the cell cycle progression after desynchronization.Methods The doubling time of HEC-1-A cells was detected by cell counting Kit-8 assay.To determine the best lovastatin concentration of G1 synchronization,HEC-1-A cells were treated with lovastatin at concentration of 10,20,30 and 40 μmol/L respectively for 1 × doubling time,and the cell cycle was detected by flow cytometry (FCM).To determine the best period of lovastatin treatment to achieve G1 synchronization,HEC-1-A cells were treated with lovastatin at the best concentration for 0.5 × to 2 × doubling time,and the cell cycle was detected every 4 h using FCM.Furthermore,the cell cycle progress of HEC-1-A cells after desynchronization was also observed.Results The doubling time of HEC-1-A cells was 24 h.Treated with lovastatin at concentration of 40 μmol/L for 28 h achieved maximum G1 arrest (87.87±0.70) % in HEC-1-A cells.Minimum G1 phase (58.42±0.54) % and maximum S phase (33.58±0.62) % were observed after desynchronizing for 20 h.Conclusion Maximum G1 synchronization of HEC-1-A cells is induced by lovastatin at concentration of 40 μmol/L for 28 h.The HEC-1-A cells show minimum G1 phase and maximum S phase after desynchronizing for 20 h.
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