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.
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50.0μmol/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、50.0μmol/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。