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目的观察不同温度下吉西他滨(GEM)热化疗抑制膀胱癌BIU-87细胞株细胞增殖和诱导凋亡的情况,并探讨其可能的机制。方法将处于对数生长期的BIU-87细胞分为4组,A组为对照组,B组为热化疗1组[(38±0.2)℃],C组为热化疗2组[(40±0.2)℃],D组为热化疗3组[(43±0.2)℃],GEM的工作浓度为1μg/ml。用四甲基偶氮唑蓝(MTT)比色法,BrdU掺入实验检测各组对细胞生长增殖的影响;流式细胞术(FCM)检测各组对细胞周期及细胞凋亡情况的影响;免疫印迹法(Western blot)检测各组细胞BcL-2,Capase-3蛋白的表达变化。结果 MTT比色法显示相对于A组,B、C、D组对BIU-87细胞的生长增殖均可产生抑制作用,其中D组的抑制作用最为明显,热化疗组与对照组之间的差异均有统计学意义(P<0.05)。BrdU掺入实验提示B、C、D组细胞处于S期的比例明显少于A组(P<0.05),其中D组的比例最低。FCM检测显示,BIU-87经处理后48 h后实验组细胞周期有明显的变化,G0/G1期细胞比例上升,S期细胞比例下降,其中以D组变化最为明显,与A组相比差异有统计学意义(P<0.05)。相对于对照组,实验组各组的凋亡率明显增高,其中以D组最为明显(P<0.05)。Western blotting结果显示,试验组的Caspase-3蛋白表达活性增高,而BcL-2蛋白表达降低,也以D组变化最为明显。结论 GEM热化疗能抑制BIU-87细胞的增殖,并通过下调BcL-2蛋白表达,上调Caspase-3蛋白表达活性,进而诱导细胞凋亡,并随着温度的升高越明显。
Objective To observe the effect of gemcitabine (GEM) thermotherapy on the proliferation and apoptosis of bladder cancer cell line BIU-87 at different temperatures and to explore its possible mechanism. Methods BIU-87 cells in logarithmic growth phase were divided into 4 groups, group A was control group, group B was thermochemotherapy group [(38 ± 0.2) ℃], group C was thermochemotherapy group (40 ± 0.2) ° C], group D was 3 groups [(43 ± 0.2) ° C], and the working concentration of GEM was 1 μg / ml. The effect of each group on cell proliferation and proliferation was detected by MTT assay and BrdU incorporation assay. The effect of each group on cell cycle and apoptosis was detected by flow cytometry (FCM) Western blot was used to detect the expression of Bcl-2 and Capase-3 in each group. Results The results of MTT assay showed that BIU-87 cells could inhibit the growth of BIU-87 cells in comparison with groups A, B, C and D, and the inhibitory effect of D group was the most obvious. The difference between the thermochemotherapy group and the control group All were statistically significant (P <0.05). BrdU incorporation experiments suggest that B, C, D cells in the S phase was significantly less than the A group (P <0.05), of which the lowest proportion of D group. The results of FCM showed that the cell cycle of experimental group changed obviously after 48 h treatment, the proportion of cells in G0 / G1 phase increased and the percentage of S phase cells decreased, especially in group D, which was significantly different from that in group A There was statistical significance (P <0.05). Compared with the control group, the apoptosis rate of the experimental group was significantly increased, especially in the D group (P <0.05). Western blotting results showed that Caspase-3 protein expression activity increased, while BcL-2 protein expression decreased, also the most obvious changes in the D group. Conclusions GEM thermo-chemotherapy can inhibit the proliferation of BIU-87 cells and up-regulate the expression of Bcl-2 protein and up-regulate the activity of Caspase-3 protein and induce the apoptosis of BIU-87 cells.