论文部分内容阅读
目的:探讨米非司酮抑制前列腺癌PC-3细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达过程中糖皮质激素受体(glucocorticoid receptor,GR)信号通路的作用。方法:采用免疫细胞化学法检测前列腺癌PC-3细胞中GR的表达情况,然后采用RT-PCR法检测不同浓度米非司酮作用48h后PC-3细胞中GR mRNA的表达变化,并用ELISA法检测不同浓度米非司酮作用不同时间以及不同激素作用后PC-3细胞中VEGF蛋白表达的变化。结果:免疫细胞化学检测证实了前列腺癌PC-3细胞中存在GR的表达。进一步用RT-PCR法检测发现,5、10和50μmol/L米非司酮作用PC-3细胞48h后GR mRNA的表达呈逐渐降低趋势,其中5μmol/L米非司酮组与未处理对照组差异无统计学意义(P>0.05),而10和50μmol/L米非司酮组与对照组比较,差异有统计学意义(P<0.01)。不同浓度的米非司酮作用PC-3细胞48h后,其VEGF的分泌量随着米非司酮浓度的增加而逐渐减少,其中≥10μmol/L的米非司酮组VEGF分泌量比对照组下降≥48.29%;而10μmol/L米非司酮作用PC-3细胞24～96h后,其VEGF的分泌量随着作用时间延长而呈递减趋势,其中48h后的VEGF分泌量比对照组下降超过48.23%,差异有统计学意义(P<0.01)。不同激素作用PC-3细胞后,除米非司酮可减少VEGF的表达(P<0.01)外,孕激素可使VEGF蛋白的表达水平略有升高,而其他激素对VEGF蛋白的表达无影响;同时,地塞米松可以部分阻断米非司酮对PC-3细胞中VEGF蛋白的抑制作用。结论:米非司酮抑制前列腺癌细胞分泌VEGF蛋白,可能是通过GR信号通路发挥作用的。
Objective: To investigate the effect of mifepristone on the signal transduction pathway of glucocorticoid receptor (GR) in the expression of vascular endothelial growth factor (VEGF) in PC-3 cells. Methods: The expression of GR in PC-3 cells was detected by immunocytochemistry. The expression of GR mRNA in PC-3 cells was detected by RT-PCR 48 h after treatment with different concentrations of mifepristone. The changes of VEGF protein expression in PC-3 cells treated with different concentrations of mifepristone at different times and different hormones were detected. Results: Immunocytochemistry confirmed the presence of GR in prostate cancer PC-3 cells. The results of RT-PCR showed that the expression of GR mRNA in PC-3 cells treated with 5, 10 and 50μmol / L mifepristone gradually decreased, and the expression of GR mRNA in PC-3 cells treated with 5μmol / L mifepristone and untreated control group (P> 0.05), while there was significant difference between 10 and 50 μmol / L mifepristone group and the control group (P <0.01). After 48h exposure to different concentrations of mifepristone, the secretion of VEGF decreased gradually with the increase of mifepristone concentration, and the secretion of VEGF in mifepristone group ≥10μmol / L was higher than that of control group Decreased 48.29%, while the secretion of VEGF in PC-3 cells treated with 10μmol / L mifepristone decreased gradually with the prolongation of action time, and the secretion of VEGF decreased more than 48h 48.23%, the difference was statistically significant (P <0.01). Progesterone increased the expression of VEGF protein in PC-3 cells except mifepristone (P <0.01), while other hormones had no effect on the expression of VEGF protein At the same time, dexamethasone partially blocked the inhibitory effect of mifepristone on VEGF protein in PC-3 cells. Conclusion: Mifepristone inhibits the secretion of VEGF protein in prostate cancer cells, which may play a role through the GR signaling pathway.