曲古抑菌素A和紫杉醇对肺腺癌细胞增殖和凋亡的影响

Effect of trichostatin A and paclitaxel on the growth and apoptosis of lung adenocarcinoma cell lines

目的研究曲古抑菌素A（TSA）联合紫杉醇对肺腺癌细胞增殖和凋亡的影响。方法以TSA和紫杉醇单独或联合处理A549细胞后，采用锥虫蓝拒染法观察药物对肿瘤细胞增殖的影响，采用Hoechst33258染色法观察细胞凋亡，采用流式细胞术检测细胞凋亡和细胞周期的变化，采用Westernblot法检测肿瘤细胞凋亡信号通路中多聚二磷酸腺苷核糖多聚酶（PARP）、casapase-3、乙酰化微管蛋白和survivin蛋白的表达变化。结果TSA和紫杉醇对A549细胞均有抑制作用，TSA联合紫杉醇抑制作用更强。250nmol／LTSA联合10μg/ml紫杉醇处理细胞72h，细胞增殖抑制率为82．8％。单用TSA或紫杉醇均可诱导A549细胞凋亡，细胞凋亡率分别为（17．6±1．8）％和（39．2±3．7）％，TSA和紫杉醇联合应用可使A549细胞的凋亡率增至（64．2±4．2）％，与对照组[（1．2±0．5）％]相比，差异有统计学意义（P〈0．01）。TSA组、紫杉醇组、TSA＋紫杉醇组G2／M期细胞百分比分别为（23．5±2．2）％、（10．5±1．5）％和（32．4±3．1）％，与对照组[（4．8±1．1）％]比较，TSA组和TSA＋紫杉醇组差异均有统计学意义（P〈0．05，P〈0．01）。紫杉醇和TSA可诱导A549细胞微管蛋白乙酰化，使survivin蛋白的表达减少，PARP和casapase-3表达增加。结论TSA和紫杉醇能抑制A549肿瘤细胞生长，诱导细胞凋亡，二者联合具有协同作用。

Objective To investigate the effect of trichostatin A （TSA）/paclitaxel on the growth and apoptosis in human lung adenocarcinoma cell line A549 cells. Methods Human lung adenocarcinoma A549 cells were cultured in DMEM in the presence of paclitaxel and the histone deacetylase inhibitor trichostatin A, and the growth curve was obtained by trypan-blue exclusion assay and cell count. Apoptosis was assessed using Hoechst 33258 staining and flow cytometry, and cell cycle was detected by flow eytometry analysis. The proteins of PARP, easpase-3, survivin and tubulin acetylation were detected by Western blotting. Results Significant growth reduction was observed in the A549 ceils following treatment with paclitaxel or the histone deacetylase inhibitor TSA. The combined treatment with TSA/paclitaxel caused the highest inhibition of cell growth. The apoptosis rate of A549 cells treated with TSA or paclitaxel for 24 hours was （ 17.6± 1.8） % and （39.2 ± 3.7） %, respectively, but a significantly higher apoptosis rate was （64.2 ±4.2） % was induced by combined treatment with TSA and paclitaxel. In contrast with the control group, the cell cycle was markedly arrested at G2/M phase in the TSA and paclitaxel group （P 〈 0.05 ）. The Western blot analysis demonstrated that treatment with TSA/paclitaxel led to a synergistic increase of acetylated tubulin, PARP and caspase-3, and reduced the expression of survivin. Conclusion TSA or paclitaxel alone can inhibit the cell growth and induce apoptosis, and the combination of TSA and paclitaxel exerts a synergistic effect on the growth and apoptosis in lung adenocarcinoma cells.