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目的评价吉西他滨(GEM)、紫杉醇(TAX)时序联合抗膀胱癌细胞系效应。方法MTT法检测GEM与TAX同时(A组)、先后间隔24h时序(B组为GEM先于TAX24h,C组为TAX先于GEM24h)联合用药对膀胱癌细胞系5637和EJ细胞的细胞毒活性,Chou和Talalay方法评估两药物作用关系;流式细胞仪碘化丙啶法检测药物作用细胞后的细胞周期分布变化;AnnexinV检测各组细胞凋亡率的变化;蛋白印迹法检测细胞周期CyclinD1和CyclinB1蛋白的变化。结果药物浓度为1.0μg/ml时,A组5637、EJ细胞生长抑制率为38%、40%,细胞凋亡率为29.3%、15.4%,细胞毒联合指数(CI)均<1;C组5637、EJ细胞生长抑制率为15%、13%,细胞凋亡率为11.5%、6.7%,CI均>1;B组结果与A组相仿。给药第48h细胞周期分布显示细胞B组主峰在G1期积聚(≥60%),A组G1、G2/M期略有增加,C组主峰在G2/M期积聚(≥65%)。A组CyclinD1蛋白表达较无药物对照组下降,CyclinB1表达上调。结论联合应用GEM和TAX治疗膀胱癌,存在时序效应,即同时用药产生协同作用,先用GEM,后用TAX也可产生较好的细胞毒效应,而先用TAX,后用GEM,则产生拮抗作用。
Objective To evaluate the effect of combination of gemcitabine (GEM) and paclitaxel (TAX) on anti-bladder cancer cell line. Methods MTT assay was used to detect the cytotoxicity of GEM combined with TAX on bladder cancer cell line 5637 and EJ cells at the interval of 24 h (group B preceded TAX 24 h and group C TAX before GEM 24 h) Chou and Talalay methods were used to evaluate the relationship between the two drugs. The change of cell cycle distribution was detected by flow cytometry with propidium iodide method. The apoptosis rate of each group was detected by AnnexinV. The expressions of CyclinD1 and CyclinB1 Changes in protein. Results When the drug concentration was 1.0μg / ml, the growth inhibition rate of EJ cells in group A was 38%, 40%, the apoptosis rate was 29.3%, 15.4%, the cytotoxicity index (CI) 5637, EJ cell growth inhibition rate was 15%, 13%, the apoptosis rate was 11.5%, 6.7%, CI were> 1; The 48h cell cycle distribution showed that the main peak of cell group B accumulated in G1 phase (≥60%), G1 and G2 / M phase increased slightly in group A, and the main peak of C group accumulated in G2 / M phase (≥65%). The expression of CyclinD1 in group A was lower than that in the non-drug control group, and the expression of CyclinB1 was up-regulated. Conclusions The combination of GEM and TAX in the treatment of bladder cancer, there is a timing effect, that is synergistic with the drug at the same time, first with GEM, followed by TAX can produce better cytotoxic effect, but first with TAX, after GEM, antagonism effect.