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目的:研究染料木黄酮(GDN)对去势抵抗性前列腺癌细胞VCa P增殖的抑制作用及可能机制。方法:以不同浓度(0、12.5、25、50、100、200μmol/L)的GEN处理去势抵抗性前列腺癌细胞VCaP,分别在24、48、72 h以CCK-8法测定细胞增殖;以流式细胞术检测细胞周期;以免疫荧光法检测Ki-67表达水平;以Western印迹法检测Cyclin D1、PCNA、P53及PSA表达水平。结果:12.5、25、50、100、200μmol/L GEN作用于VCaP细胞72 h后,对VCaP细胞的抑制率分别为(25.38±0.02)%、(31.14±0.29)%、(45.27±0.03)%、(52.19±0.05)%与(68.21±0.19)%,与对照组[(10.08±0.02)%]相比差异有统计学意义(P<0.05)。流式细胞术显示GEN可导致VCa P细胞G2/M期阻滞(P<0.05)。免疫细胞化学法检测显示GEN能够抑制细胞中Ki-67的表达。Western印迹显示去势抵抗性前列腺癌细胞内PSA、Cyclin D1、PCNA随着GEN浓度的增加逐渐下调,P53随着GEN浓度的增加逐渐上调(P<0.05)。结论:GEN能够抑制体外培养的去势抵抗性前列腺癌细胞VCaP增殖,其作用机制可能与阻滞细胞周期并伴随相关周期蛋白表达的改变有关。
AIM: To investigate the inhibitory effect of genistein (GDN) on VCa P proliferation in ovariectomized prostate cancer cells and its possible mechanism. Methods: The castration-resistant prostate cancer cells VCaP were treated with different concentrations of GEN (0, 12.5, 25, 50, 100 and 200μmol / L). Cell proliferation was measured by CCK-8 at 24, The cell cycle was detected by flow cytometry. Ki-67 expression was detected by immunofluorescence. The expression of Cyclin D1, PCNA, P53 and PSA were detected by Western blotting. Results: The VCaP cells treated with 12.5, 25, 50, 100 and 200 μmol / L GEN for 72 h inhibited the proliferation of VCaP cells by (25.38 ± 0.02)%, (31.14 ± 0.29)%, (45.27 ± 0.03)%, , (52.19 ± 0.05)% and (68.21 ± 0.19)%, respectively, compared with that in the control group [(10.08 ± 0.02)%] (P <0.05). Flow cytometry showed that GEN could lead to G2 / M arrest of VCa P cells (P <0.05). Immunocytochemistry showed that GEN can inhibit the expression of Ki-67 in cells. Western blotting showed that the levels of PSA, Cyclin D1 and PCNA in ovariectomized prostate cancer cells decreased gradually with the increase of GEN concentration, while P53 increased gradually with the increase of GEN concentration (P <0.05). CONCLUSION: GEN can inhibit VCaP proliferation in ovariectomized prostate cancer cells in vitro. Its mechanism may be related to arresting cell cycle and accompanying changes of related cyclin expression.