曲古抑菌素A联合多西他赛促进肺腺癌细胞的凋亡及其可能的分子机制

Trichostatin A combined with docetaxel promote apoptosis of lung adenocarcinoma cells and the possible molecular mechanisms

目的:研究曲古抑菌素A(trichostatin A,TSA)联合多西他赛(docetaxel,Doc)对肺腺癌细胞A549凋亡的影响及其分子机制。方法:TSA单独或联合Doc处理A549细胞,MTT法检测A549细胞的增殖,光学显微镜下观察A549细胞的形态学变化,Hoechst 33258染色及流式细胞术检测A549细胞的凋亡,流式细胞术检测细胞周期的变化,Western blotting检测乙酰化-α-微管蛋白(acetyl-α-tubulin)、survivin蛋白的表达及caspase-3的活化。结果:10μg/ml Doc、250 nmol/L TSA单独或联合作用均能时间依赖性地抑制A549细胞的增殖,联合作用的抑制率显著高于Doc或TSA单独作用[(65.6±3.1)%vs(30.6±2.1)%、(23.3±1.9)%,P<0.05]。TSA、Doc单独或联合用药均可使A549细胞凋亡率增加,联合作用的凋亡率显著高于单独作用[(58±3.6)%vs(17±2.2)%、(14±1.6)%,P<0.05]。联合作用比单独作用使A549细胞更明显地阻滞于G2/M期[(32.4±3.1)%vs(23.5±2.3)%、(10.5±1.5)%,P<0.05]。TSA联合Doc可进一步增加acetyl-α-tubulin表达、减少sur-vivin蛋白的表达,并促进casapase-3的活化(均P<0.05)。结论:TSA联合Doc能够抑制A549细胞的增殖、促进细胞的凋亡,其机制可能与上调acetyl-α-tubulin的表达、抑制survivin的表达以及促进caspase-3活化有关。

Objective：To explore the effects of trichostatin A（TSA） and docetaxel（Doc） on apoptosis of lung adenocarcinoma A549 cells and the possible molecular mechanisms.Methods： A549 cells were treated with TSA alone or in combination with Doc,MTT assay was used to measure the proliferation of A549 cells;cell morphological changes were observed under light microscope;apoptosis was assessed using Hoechst 33258 staining and flow cytometry;cell cycle was analyzed by flow cytometry;the expression of acetyl-α-tubulin and survivin protein and activation of caspase-3 were detected by Western blotting.Results： 10 μg/ml Doc and 250 nmol/L TSA alone or in combination significantly inhibited the growth of A549 cells in a time-dependent manner,and the combined treatment induced even higher inhibitory rate（[65.6±3.1]% vs [30.6±2.1]%,[23.3±1.9]%,P0.05）.In addition,TSA or Doc alone or in combination increased the apoptosis rate of A549 cells,and the combined treatment also induced higher apoptosis rate（[58±3.6]% vs [17±2.2]%,[14±1.6]%,P0.05）.The cell cycle was more markedly arrested in G2/M phase in combination treatment group（[32.4±3.1]% vs [23.5±2.3]%,[10.5±1.5]%,P0.05）,TSA combined with Doc increased the expression of acetyl-α-tubulin,reduced the expression of survivin and promoted the activation of caspase-3（P0.05）.Conclusion： TSA combined with Doc can inhibit proliferation and induce apoptosis of A549 cells,which is related to the up-regulation of acetyl-α-tubulin,down-regulation of survivin and increased activation of caspase-3.