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目的:研究曲古抑菌素A(trichostatin A,TSA)和紫杉醇(paclitaxel,PTX)对人子宫内膜癌细胞株KLE增殖和凋亡的影响。方法:TSA和PTX、卡铂(carboplatin,Carbo)、多柔比星(doxorubicin,Dox)单独或联合作用于KLE细胞,锥虫蓝法观察药物对肿瘤细胞生长的影响;Annexin V、Hoechst染色和线粒体膜电位检测细胞凋亡;Western blotting检测肿瘤细胞凋亡信号通路中多聚ADP核糖聚合酶(PARP)、半胱天冬氨酸蛋白酶9(caspase-9)和乙酰化微管蛋白的表达。结果:PTX、Carbo、Dox和TSA对KLE细胞增殖均有抑制作用,TSA和PTX联用后抑制作用最强。Annexin V染色、Hoechst染色、线粒体膜电位法和PARP、Caspase-9的检测显示,单用PTX或TSA均可诱导细胞凋亡,联合应用产生最强的协同作用。Western blotting和免疫组化分析显示,PTX和TSA均可诱导微管蛋白乙酰化,联合用药后微管蛋白乙酰化明显增加。结论:TSA和PTX联合使用有明显的协同作用,能显著抑制子宫内膜癌KLE细胞生长和诱导细胞凋亡,其机制与激活线粒体凋亡信号通路和增强微管蛋白乙酰化有关。
Objective: To study the effects of trichostatin A (TSA) and paclitaxel (PTX) on the proliferation and apoptosis of human endometrial carcinoma cell line KLE. Methods: TSA and PTX, carboplatin (Carbo) and doxorubicin (Dox) alone or in combination were applied to KLE cells. The effects of drugs on the growth of tumor cells were observed by trypan blue method. Annexin V, Hoechst staining and Mitochondrial membrane potential was used to detect apoptosis. Western blotting was used to detect the expression of poly ADP ribose polymerase (PARP), caspase-9 and acetylated tubulin in apoptotic signaling pathway. Results: PTX, Carbo, Dox and TSA inhibited the proliferation of KLE cells, and the combination of TSA and PTX had the strongest inhibitory effect. Annexin V staining, Hoechst staining, mitochondrial membrane potential and PARP, Caspase-9 test showed that the single use of PTX or TSA can induce apoptosis, combined application of the strongest synergistic effect. Western blotting and immunohistochemical analysis showed that both PTX and TSA induced tubulin acetylation, and tubulin acetylation was significantly increased after combined administration. CONCLUSION: The combination of TSA and PTX has obvious synergistic effect, which can significantly inhibit the growth of KLE cells and induce apoptosis of endometrial carcinoma cells. The mechanism is related to activation of mitochondrial apoptosis signaling pathway and enhancement of tubulin acetylation.