【摘要】 目的探讨热休克蛋白90(HSP90)抑制剂17-丙烯胺基-17-去甲氧格尔德霉素(17-AAG)联合紫杉醇对人未分化甲状腺癌FRO细胞增殖和凋亡的影响。方法 1采用四甲基偶氮唑盐微量酶反应比色法(MTT比色法)测定不同浓度(17-AAG:0.312 5、0.625 0、1.250 0、2.500 0及5.000 0μmol/L;紫杉醇:0.001 0、0.010 0、0.100 0及1.000 0μmol/L)、不同时间(24、48及72 h)17-AAG、紫杉醇单药和联合处理(17-AAG:0.625 0μmol/L,紫杉醇:0.001 0、0.010 0、0.100 0及1.000 0μmol/L)后FRO细胞的增殖抑制率。2采用流式细胞仪检测17-AAG、紫杉醇单药及联合处理24 h后(17-AAG:0.625 0μmol/L、紫杉醇:0.100 0μmol/L;联合用药:17-AAG的浓度为0.625 0μmol/L,紫杉醇的浓度为0.100 0μmol/L)FRO细胞的细胞周期变化及凋亡率。3采用胱天蛋白酶-3(Caspase-3)和Caspase-9检测试剂盒检测17-AAG、紫杉醇单药及联合处理24 h后(17-AAG:0.625 0μmol/L、紫杉醇:0.100 0μmol/L;联合用药:17-AAG的浓度为0.625 0μmol/L,紫杉醇的浓度为0.100 0μmol/L)后FRO细胞中的Caspase-3和Caspase-9活性。空白对照组均不加任何药物,只加培养液。结果 1同时点空白对照组、各剂量17-AAG组/紫杉醇组/17-AAG联合紫杉醇组的增殖抑制率随浓度升高而逐渐升高,任2组比较差异均有统计学意义(P<0.05);各剂量17-AAG组/紫杉醇组/17-AAG联合紫杉醇组的增殖抑制率在24、28及72 h逐渐增高,任2组比较差异均有统计学意义(P<0.05);同时点同浓度情况下,17-AAG联合紫杉醇组的增殖抑制率均高于单独用药组(P<0.05)。各时点17-AAG与紫杉醇联合的q值均大于1.15,两者之间呈协同作用。2 17-AAG组、紫杉醇组及17-AAG联合紫杉醇组FRO细胞的凋亡率均明显高于空白对照组(P<0.05),且17-AAG联合紫杉醇组FRO细胞的凋亡率高于17-AAG组和紫杉醇组(P<0.05)。3 17-AAG组、紫杉醇组及17-AAG联合紫杉醇组FRO细胞的Caspase-3和Caspase-9活性均高于空白对照组(P<0.05);且17-AAG联合紫杉醇组细胞的Caspase-3和Caspase-9活性均高于17-AAG组和紫杉醇组相应指标(P<0.05)。结论 17-AAG和紫杉醇均可明显抑制FRO细胞的增殖并诱导细胞凋亡,联合用药有一定的协同效应,呈剂量依赖关系。更多还原

【Abstract】 Objective To investigate the inhibitory effect of heat shock protein 90(HSP90) inhibitors of 17-propylene amino-17-demethoxy geldanamycin(17-AAG) combining with paclitaxel on human anaplastic thyroid cancer FRO cell line. Methods 1 The proliferation inhibition rates of FRO cells were detected by mmethyl thiazolyl tetrazolium(MTT) assay in different concentration groups(17-AAG: 0.312 5, 0.625 0, 1.250 0, 2.500 0, and 5.000 0 μmol/L; paclitaxel: 0.001 0, 0.010 0, 0.100 0, and 1.000 0 μmol/L; combination group, 17-AAG: 0.625 0 μmol/L, paclitaxel: 0.001 0, 0.010 0, 0.100 0, and 1.000 0 μmol/L) and at different time points(24, 48, and 72 hours). 2 The change of cell cycle and apoptosis rates of FRO cells were detected in 17-AAG group(0.625 0 μmol/L), paclitaxel group(0.100 0 μmol/L), and combination group(17-AAG: 0.625 0 μmol/L, paclitaxel: 0.100 0 μmol/L) by flow cytometry at 24 hours after treatment. 3 Activity of Caspase-3 and Caspase-9 in FRO cells of 17-AAG group(0.625 0 μmol/L), paclitaxel group(0.100 0 μmol/L),and combination group(17-AAG: 0.625 0 μmol/L, paclitaxel: 0.100 0 μmol/L) was detected by Caspase-3 detection reagent box and Caspase-9 detection reagent box respectively. FRO cells of normal control group were treated without any drug,but culture solution. Results 1 The proliferation inhibition rates of FRO cells increased with the increase of concentration(17-AAG, paclitaxel, combination of 17-AAG and paclitaxel), there was significant difference between any 2 groups(normal control group included), P<0.05. In addition, the proliferation inhibition rates of FRO cells in any concentration group(normal control group excluded) increased over time(24, 48, and 72 hours), there was significant difference between any 2 time points(P<0.05). The proliferation inhibition rates of FRO cells in combination group were all higher than those of 17-AAG group and paclitaxel group in condition of same time point and same concentration(P<0.05). The q value of combination group was higher than 1.15 at 3 time points in all concentration, that meant 17-AAG could increase the efficiency of paclitaxel. 2 The apoptosis rate of FRO cells in normal control group was lower than those of 17-AAG group, paclitaxel group, and combination group(P<0.05), and apoptosis rate of FRO cells in combination group was higher than those of 17-AAG group and paclitaxel group(P<0.05). 3 Activity of Caspase-3 and Caspase-9 of FRO cells in normal control group were lower than those of 17-AAG group, paclitaxel group, and combination group(P<0.05), and activity of Caspase-3 and Caspase-9 of FRO cells in combination group were higher than those of 17-AAG group and paclitaxel group(P<0.05). Conclusions 17-AAG and paclitaxel can significantly inhibit the proliferation and induce the apoptosis of FRO cells. The combination of the two kinds of drugs may generate synergy, with dose-dependence effect.更多还原