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目的:探讨曲格列酮对人前列腺癌PC-3细胞增殖、侵袭和迁移的影响。方法:PC-3细胞经不同浓度曲格列酮(0、1×10-6、1×10-5和1×10-4 mol/L)处理后,采用MTT法检测各组细胞增殖水平;细胞划痕实验和细胞侵袭试验检测侵袭及迁移能力的改变;蛋白质印迹法检测细胞Id1、MMP-2和MMP-9蛋白的表达水平。结果:MTT检测显示,不同浓度曲格列酮对PC-3细胞抑制作用明显,并呈浓度和时间依赖性,P<0.001。细胞划痕实验显示,PC-3细胞经不同浓度曲格列酮作用后,1×10-6 mol/L组细胞迁移率为(70.67±10.42)%,1×10-5 mol/L组为(49.00±6.00)%,1×10-4 mol/L组为(20.33±5.05)%,明显低于对照组的(80.5±6.86)%,差异有统计学意义,P<0.01,且呈显著剂量依赖关系,F=78.57,P<0.001。细胞体外侵袭实验显示,1×10-6 mol/L组穿越Transwell滤膜的PC-3细胞数为85.17±15.28,1×10-5 mol/L组为70.33±7.63,1×10-4 mol/L组为52.50±8.87,明显低于对照组的137.67±15.09,差异有统计学意义,P<0.001,且呈显著剂量依赖关系,F=53.99,P<0.001。细胞体外迁移实验显示,1×10-6 mol/L组迁移至Transwell下室的细胞数为88.83±17.02,1×10-5 mol/L组为71.67±9.44,1×10-4 mol/L组为54.50±9.35,明显低于对照组的142.00±17.40,差异有统计学意义,P<0.001,且呈显著剂量依赖关系,F=44.72,P<0.001。蛋白质印迹法检测结果显示,对照组Id1蛋白相对表达量为12.67±7.23,1×10-6 mol/L组为24.67±6.51,1×10-5 mol/L组为36.67±6.11,1×10-4 mol/L组为49.33±4.93;对照组MMP-2蛋白相对表达量为9.00±4.36,1×10-6 mol/L组为25.33±8.62,1×10-5 mol/L组为42.00±7.55,1×10-4 mol/L组为58.67±12.06;对照组MMP-9蛋白相对表达量为13.00±3.61,1×10-6 mol/L组为25.67±6.43,1×10-5 mol/L组为38.33±8.96,1×10-4 mol/L组为53.67±6.43。曲格列酮显著下调PC-3细胞Id1、MMP-2和MMP-9蛋白表达水平,P值均<0.05。结论:曲格列酮能有效抑制PC-3细胞增殖与侵袭迁移,同时下调Id1、MMP-2和MMP-9蛋白表达水平。
Objective: To investigate the effect of troglitazone on proliferation, invasion and migration of human prostate cancer PC-3 cells. METHODS: PC-3 cells were treated with troglitazone (0, 1 × 10-6, 1 × 10-5, and 1 × 10-4 mol / L) at different concentrations. The proliferation of PC-3 cells was detected by MTT assay. Cell scratch assay and cell invasion assay were used to detect the changes of invasion and migration ability. Western blotting was used to detect the expression of Id1, MMP-2 and MMP-9. Results: MTT assay showed that troglitazone inhibited the proliferation of PC-3 cells in a dose-dependent and time-dependent manner (P <0.001). Cell scratch assay showed that the cell migration rate of PC-3 cells treated with troglitazone was (70.67 ± 10.42)% and 1 × 10-5 mol / L (49.00 ± 6.00)% in the control group and (20.33 ± 5.05)% in the 1 × 10-4 mol / L group, which was significantly lower than that in the control group (80.5 ± 6.86)%, P <0.01 Dose-dependent, F = 78.57, P <0.001. The in vitro cell invasion assay showed that the number of PC-3 cells passing through Transwell membrane filter was 85.17 ± 15.28 in 1 × 10-6 mol / L group, 70.33 ± 7.63 and 1 × 10-4 mol in 1 × 10-5 mol / L group / L group was 52.50 ± 8.87, significantly lower than the control group 137.67 ± 15.09, the difference was statistically significant, P <0.001, and a significant dose-dependent relationship, F = 53.99, P <0.001. In vitro migration experiments showed that the number of cells migrating to the lower chamber of Transwell in the concentration of 1 × 10-6 mol / L was 88.83 ± 17.02, that in the group of 1 × 10-5 mol / L was 71.67 ± 9.44,1 × 10-4 mol / L Group was 54.50 ± 9.35, significantly lower than the control group 142.00 ± 17.40, the difference was statistically significant, P <0.001, and a significant dose-dependent relationship, F = 44.72, P <0.001. The results of Western blotting showed that the relative expression of Id1 protein in control group was 12.67 ± 7.23, that in group of 1 × 10-6 mol / L was 24.67 ± 6.51 and that in group of 1 × 10-5 mol / L was 36.67 ± 6.11,1 × 10 -4 mol / L group was 49.33 ± 4.93; the relative expression of MMP-2 protein in control group was 9.00 ± 4.36, in group of 1 × 10-6 mol / L was 25.33 ± 8.62, in group of 1 × 10-5 mol / L was 42.00 ± 7.55,1 × 10-4 mol / L group was 58.67 ± 12.06. The relative expression of MMP-9 protein in the control group was 13.00 ± 3.61 and that in the 10 × 10-6 mol / L group was 25.67 ± 6.43,1 × 10-5 mol / L group was 38.33 ± 8.96,1 × 10-4 mol / L group was 53.67 ± 6.43. Troglitazone significantly down-regulated the expression of Id1, MMP-2 and MMP-9 in PC-3 cells, both P <0.05. Conclusion: Troglitazone can effectively inhibit the proliferation, invasion and migration of PC-3 cells and down-regulate the expression of Id1, MMP-2 and MMP-9.