南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (10): 1409-.

• •    下一篇

Histone deacetylase inhibitors induce human renal cell carcinoma cell apoptosis through p-JNK activation

徐米清，洪明，谢辉   

  1. 广州医科大学附属第二医院内科，广东广州510260
  • 出版日期:2013-10-20 发布日期:2013-10-20

组蛋白去乙酰化酶抑制剂通过激活p-JNK信号通路诱导肾癌细胞凋亡

  1. 广州医科大学附属第二医院内科，广东广州510260
  • Online:2013-10-20 Published:2013-10-20

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摘要： 目的研究组蛋白去乙酰化酶抑制剂曲古抑菌素A（trichostatinA, TSA）和帕比司他（panobinostat, LBH589）在体外对人肾
癌细胞株增殖、细胞周期分布及凋亡的影响，并进一步探讨其可能的分子机制。方法LBH589 或TSA 作用于经或未经
SP600125预处理的人肾癌细胞株OS-RC-2，在设定的时间点用四甲基偶氮唑蓝方法（MTT）检测细胞生长抑制率并检测出最佳
刺激浓度和时间；用流式细胞术检测药物作用后肾癌细胞周期变化情况及凋亡情况；用Western blotting分析药物作用后肾癌细
胞内c-Jun、磷酸化c-Jun（p-c-Jun）、Bcl-2、Bax蛋白的表达变化。用JNK抑制剂SP600125阻断JNK信号通路后，观察SP600125
预处理+TSA组较TSA单独处理肾癌细胞周期、凋亡及凋亡相关蛋白的变化情况。结果TSA和LBH589在体外均可抑制肾癌
细胞生长，且具有浓度与时间依赖性；与对照组相比，TSA 或LBH589 可使肾癌细胞发生G2/M期周期阻滞，并均可诱导肾癌细
胞发生明显凋亡。Western blotting显示TSA和LBH589均能明显促进p-c-jun蛋白的表达，同时TSA也能诱导Bax蛋白表达而
抑制Bcl2 蛋白表达，与对照组相比有显著统计学意义（P<0.05）；与TSA单独处理相比，JNK抑制剂SP600125预处理+TSA能部
分减弱TSA对肾癌细胞的抑制效应并逆转TSA诱导的肾癌细胞G2/M期阻滞和细胞凋亡，差异有统计学意义（P<0.05）。结论
组蛋白去乙酰化酶抑制剂在体外可抑制OS-RC-2 细胞增殖，阻滞细胞周期、诱导细胞凋亡；TSA可通过JNK信号通路诱导
OS-RC-2细胞G2/M期阻滞，调节凋亡蛋白的表达从而诱导细胞凋亡。

Abstract: Objective To study the effect of histone deacetylase inhibitors trichostatin A (TSA) and LBH589 on the growth of
human renal cell carcinoma OS-RC-2 cells in vitro and explore the underlying molecular mechanism. Methods OS-RC-2 cells
were treated with LBH589 or TSA with or without SP600125 pretreatment, and the cell viability was measured by MTT assay.
The changes of cell cycle distribution and apoptosis of OS-RC-2 cells were examined by flow cytometry, and the expressions of
c-Jun, p-c-Jun, Bcl-2, and Bax were quantified by Western blotting. Results TSA and LBH589 both inhibited the growth of
OS-RC-2 cells in a dose- and time-dependent manner. TSA at 1 μnmol/L and LBH589 at 50 nmol/L caused obvious cell cycle
arrest in G2/M phase and cell apoptosis, and significantly increased the protein levels of phosphorylated c-Jun. TSA treatment
obviously increased Bax expression but decreased Bcl2 expression in the cells. The growth inhibitory effect of TSA was
attenuated by the JNK inhibitor SP600125 in OS-RC-2 cells. TSA-induced phosphorylation of c-Jun and Bax upregulation was
partially counteracted by SP600125. Conclusion TSA and LBH589 can cause cell cycle arrest and induce apoptosis in OS-RC-2
cells, in which process P-JNK pathway plays an important role.