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Journal of ZheJiang University(Medical Science)  2016, Vol. 45 Issue (4): 356-363    DOI: 10.3785/j.issn.1008-9292.2016.07.05
Effect of a novel EZH2 inhibitor GSK126 on prostate cancer cells
LIN Weiren1, CHEN Yatian2, ZENG Linghui2, YING Rongbiao1, ZHU Feng2
1. Department of Internal Medicine, Taizhou Cancer Hospital, Wenling 317502, China;
2. School of Medicine, Zhejiang University City College, Hangzhou 310015, China
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Objective: To investigate the effect of a novel EZH2 inhibitor GSK126 on cell growth, apoptosis and migration of prostate cancer cells. Methods: Prostate cancer PC-3 and DU145 cells were treated with GSK126 at different doses. Cell growth was detected by sulforhodamine assay. Cell apoptosis was assayed by Annexin V-/PI kit. Transwell chamber and wound healing assays were conducted to detect cell migration. The mRNA level was detected by quantitative PCR, and protein expression was detected by Western blot analysis. Results: GSK126 showed significant effect on cell growth and apoptosis when the dose was higher than 50 μmol/L. Wound healing assay revealed that scratch space in PC-3 cells was significantly increased in a dose-dependent manner in GSK126-treated groups[(247.2±24.4),(347.2±19.2) and (410.5±18.1) μm in low, medium and high dose (5.0, 20.0, 50.0 μmol/L), respectively] as compared with the control group[(171.3±17.8) μm](all P<0.05). Transwell assay showed that migrated PC-3 cells in control group was 322.0±17.9,while those in GSK126-treated groups were 198.3±15.4 (low),82.7±6.2 (medium) and 30.2±4.1 (high), and the differences between the control group and GSK126-treated groups were significant(all P<0.05). In addition, GSK126 up-regulated E-cadherin mRNA expression and down-regulated N-cadherin and Vimentin mRNA expression, whereas had no significant effect on Snail, Fibronectin and VEGF-A mRNA expression. The protein expression of E-cadherin was elevated but VEGF-A protein did not change in GSK126-treated groups. Similar results were exhibited in DU145 cell. Conclusion: GSK126 can significantly inhibit cell migration and invasion in prostate cancer PC-3 and DU145 cells, which may be resulted from its effect on epithelial-mesenchymal transition. GSK126 may be used as a potential anti-prostate cancer dug in clinic.

Key wordsGenes, tumor suppressor/drug effects      Prostatic neoplasms/drug therapy      Neoplasm proteins/biosynthesis      Protein methyltransferases      Cell movement      Cell proliferation      Apoptosis      Cell line, tumor     
Received: 18 February 2016     
CLC:  R73-3  
Cite this article:

LIN Weiren, CHEN Yatian, ZENG Linghui, YING Rongbiao, ZHU Feng. Effect of a novel EZH2 inhibitor GSK126 on prostate cancer cells. Journal of ZheJiang University(Medical Science), 2016, 45(4): 356-363.

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目的:探究zeste基因增强子同源物2(EZH2)抑制剂GSK126对前列腺癌细胞增殖、凋亡及迁移的作用及机制。方法:以磺酰罗丹明B实验考察GSK126对PC-3和DU145细胞增殖的影响,以Annexin V/PI双染法考察GSK126对细胞凋亡的影响,以Transwell实验和细胞划痕实验观察GSK126对细胞迁移和侵袭的影响,以荧光定量PCR检测GSK126对肿瘤相关基因mRNA表达的影响,以蛋白质印迹法检测相关蛋白水平的变化。结果:GSK126仅在50.0 μmol/L的高浓度剂量时能有效抑制PC-3和DU145细胞增殖,并促进其凋亡。经GSK126小、中、大剂量(5.0、20.0、50.0 μmol/L)处理后,PC-3细胞划痕间距分别为(247.2±24.4)μm、(347.2±19.2)μm、(410.5±18.1)μm,与对照组(171.3±17.8)μm比较差异均有统计学意义(均P<0.05),且呈一定的量效关系。Transwell实验结果显示,每视野下侵袭的PC-3细胞数对照组为322.0±17.9,而经GSK126小、中、大剂量处理后分别为198.3±15.4、82.7±6.2、30.2±4.1,与对照组比较差异均有统计学意义(均P<0.05)。GSK126处理前列腺癌细胞后,与上皮细胞间质转化相关的E-cadherin基因mRNA表达量上升,N-cadherin、Vimentin基因mRNA表达量下降,但Snail、Fibronectin、VEGF-A基因的mRNA表达无变化。同时,蛋白质印迹法检测结果提示,E-cadherin的蛋白表达量增加,而VEGF-A蛋白表达量无明显改变。DU145细胞上述实验结果相似。结论:GSK126能有效抑制PC-3细胞和DU145细胞迁移和侵袭,其机制与抑制上皮细胞间质转化相关。GSK126可作为潜在的前列腺癌临床治疗用药。

关键词: 基因,肿瘤抑制/药物作用,  前列腺肿瘤/药物疗法,  肿瘤蛋白质类/生物合成,  蛋白甲基转移酶类,  细胞运动,  细胞增殖,  细胞凋亡,  细胞系,肿瘤 
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