曲古抑菌素A和紫杉醇对子宫内膜癌细胞凋亡及微管稳定的影响：一项体外研究

[Effects of trichostatin A and paclitaxel on apoptosis and microtubule stabilization in endometrial carcinoma cells: an in vitro research].

作者信息

Jiang Shu-Juan, Zhang Song, Mu Xiao-Yan, Li Wei, Wang Yan

机构信息

Department of Respiratory Medicine, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2008 Sep 9;88(34):2427-31.

PMID:19087722

Abstract

OBJECTIVE

To investigate the effects of trichostatin A (TSA) and paclitaxel (PTX) on the apoptosis and microtubulin stabilization in human endometrial carcinoma cells and its mechanism.

METHODS

Human endometrial carcinoma cells of the line Ark2, KLE and AN3 were cultured in the presence of TSA (TSA group), or PTX (PTX group), or TSA plus PTX (TSA + PTX group) respectively. The growth curve was obtained by trypan-blue exclusion assay. Apoptosis was observed by annexin V and Hoechst staining. Perturbation of mitochondrial membrane potential (MMP) was detected by flow cytometry. Western blotting was used to detect the protein expression of caspase-9, poly ADP-ribose polymerase (PARP), and acetylated microtubulin.

RESULTS

The growth of the Ark2, KLE, and AN3 cells of the TSA, PTX, and TSA + PTX group, especially in the latter group, was inhibited. The Ark2 cell apoptotic rates 4 days later of the TSA, PTX, TSA + PTX, and control group were 4.25% +/- 0.25%, 12.12% +/- 0.62%, 16.56% +/- 0.74%, and 46.78% +/- 2.68% respectively by annexin V staining, and 3.39% +/- 0.12%, 6.00% +/- 0.25%, 10.05% +/- 0.53%, and 22.30% +/- 1.25% respectively by Hoechst staining. The apoptotic rates of the TSA + PTX group by both staining methods were both significantly higher than those of the other groups (all P < 0.05). Lysis of caspase-9 and PARP in the Ark2 and KLE cells increased greatly 24 hours after the TSA and PTX treatment. The disappearance rate of MMP in the Ark2 and AN3 cells of the TSA + PTX groups were 16.80% +/- 0.92% and 11.28% +/- 0.78% respectively, significantly higher than that of the PTX group (5.34% +/- 0.45% and 5.61% +/- 0.56% respectively) and TSA group (4.96% +/- 0.47% and 6.46% +/- 0.62% respectively, all P < 0.05). The expression of acetylated microtubulin was increased in the Ark2 and KLE cells of the TSA + PTX groups.

CONCLUSIONS

Synergy of TSA and PTX inhibits the cell growth and induces apoptosis. The acetylation of non-histone protein induced by histone deacetylase inhibitor is one of the possible mechanisms of its anti-cancer effects.

摘要

目的

探讨曲古抑菌素A（TSA）和紫杉醇（PTX）对人子宫内膜癌细胞凋亡及微管蛋白稳定性的影响及其机制。

方法

分别用TSA（TSA组）、PTX（PTX组）、TSA加PTX（TSA + PTX组）培养人子宫内膜癌Ark2、KLE和AN3细胞系。通过台盼蓝排斥试验获得生长曲线。采用膜联蛋白V和Hoechst染色观察细胞凋亡。用流式细胞术检测线粒体膜电位（MMP）的扰动。采用蛋白质免疫印迹法检测半胱天冬酶-9、聚ADP核糖聚合酶（PARP）和乙酰化微管蛋白的蛋白表达。

结果

TSA组、PTX组和TSA + PTX组的Ark2、KLE和AN3细胞生长受到抑制，尤其是后一组。TSA组、PTX组、TSA + PTX组和对照组4天后Ark2细胞的凋亡率，通过膜联蛋白V染色分别为4.25%±0.25%、12.12%±0.62%、16.56%±0.74%和46.78%±2.68%，通过Hoechst染色分别为3.39%±0.12%、6.00%±0.25%、10.05%±0.53%和22.30%±1.25%。两种染色方法检测TSA + PTX组的凋亡率均显著高于其他组（均P < 0.05）。TSA和PTX处理24小时后，Ark2和KLE细胞中半胱天冬酶-9和PARP的裂解显著增加。TSA + PTX组的Ark2和AN3细胞中MMP的消失率分别为16.80%±0.92%和11.28%±0.78%，显著高于PTX组（分别为5.34%±0.45%和5.61%±0.56%）和TSA组（分别为4.96%±0.47%和6.46%±0.62%，均P < 0.05）。TSA + PTX组的Ark2和KLE细胞中乙酰化微管蛋白的表达增加。