孕激素通过FOXO1对卵巢癌细胞增殖、迁移和侵袭的影响及机制

The effect and mechanism of progesterone on proliferation,migration and invasion of ovarian cancer cells through FOXO1

目的探索孕激素(Progesterone,PROG)及FOXO1抑制剂AS1842856(AS)对卵巢癌HO-8910细胞增殖、迁移和侵袭的影响及其机制。方法先用CCK-8法检测不同浓度的孕激素(0、2.5、5、10、20μmol/L)和AS(0、0.25、0.5、1、2、4μmol/L)分别作用于HO-8910细胞24、48、72 h后的增殖情况,再将HO-8910细胞分为对照组、孕激素(10μmol/L)组、AS(1μmol/L)组、AS(1μmol/L)+孕激素(10μmol/L)组4组,作用48 h,分别用CCK-8法检测各组细胞的增殖情况;Transwell迁移和侵袭实验检测各组细胞的迁移和侵袭情况;Western Blot检测各组细胞的FOXO1和β-catenin蛋白表达情况。结果不同浓度的孕激素和不同浓度的AS分别作用于HO-8910细胞不同时间后:(1)与对照组相比,除2.5μmol/L孕激素作用24 h组无显著差异外,其余各组均显示抑制作用。孕激素浓度越高,作用时间越长,对HO-8910细胞增殖的抑制作用越强。(2)与对照组相比,除4μmol/LAS作用48 h,2μmol/L AS作用72 h,4μmol/L AS作用72 h时表现为抑制作用外,其余不同作用时间不同浓度组的AS对HO-8910细胞均具有促进作用,且在AS浓度为1μmol/L时,促进作用最强。分别比较各组HO-8910细胞的增殖、迁移和侵袭能力结果:与对照组相比,孕激素组的增殖、迁移和侵袭能力均降低,AS组的增殖、迁移和侵袭能力均增高,AS+孕激素组的增殖、迁移和侵袭能力均降低。与孕激素组相比,AS+孕激素组的增殖、迁移和侵袭能力均增高。与AS组相比,AS+孕激素组的增殖、迁移和侵袭能力均降低。分别比较各组HO-8910细胞的蛋白表达结果:与对照组相比,孕激素组的FOXO1蛋白表达增多,β-catenin蛋白表达减少;AS组的FOXO1蛋白表达减少,β-catenin蛋白表达增多;AS+孕激素组的FOXO1蛋白表达减少,β-catenin蛋白表达减少。与孕激素组相比,AS+孕激素组的FOXO1蛋白表达减少,β-catenin蛋白表达增多。与AS组相比,AS+孕激素组的FOXO1蛋白表达无统计学差异,β-catenin蛋白表达减少。结论孕激素对卵巢癌HO-8910细胞的增殖、迁移和侵袭均具有抑制作用,该作用可能通过促进FOXO1进而抑制β-catenin的蛋白表达来实现。

Objective To investigate the effects and related mechanism of progesterone and FOXO1 inhibitor AS1842856(AS)on proliferation,migration and invasion of ovarian cancer HO-8910 cells.Methods First,CCK-8 method was used to detect the effects of different concentrations of progesterone(0,2.5,5,10,20μmol/L)and AS(0,0.25,0.5,1,2,4μmol/L)on the proliferation of HO-8910 cells after 24,48 and 72 h,respectively.Then,HO-8910 cells were divided into control group,progesterone(10μmol/L)group,AS(1μmol/L)group,AS(1μmol/L)+progesterone(10μmol/L)group for 48 h.The proliferation of cells in each group was detected by CCK-8 method.The migration and invasion of cells in each group were detected by Transwell migration and invasion assay.The expression levels of FOXO1 andβ-catenin of cells in each group were detected by Western blot assay.Results 1.After HO-8910 cells were treated with different concentrations of progesterone and AS in different time periods,results showed that:Compared with the control group,except the group treated with 2.5μmol/L progesterone for 24 h had no significant difference,the other groups showed inhibitory effects.The higher the concentration of progesterone,the longer the action time,and the stronger the inhibition of cell proliferation was shown.Compared with the control group,AS in the groups of different time and concentration had promoting effect on HO-8910 cells,except 4μmol/L AS for 48 h,2μmol/L AS for 72 h,and 4μmol/L AS for 72 h showed inhibitory effects.When the concentration of AS was 1μmol/L,the promoting effect was the strongest.2.(1)The cell proliferation,migration and invasion ability of each group were compared,respectively.Compared with the control group,the cell proliferation,migration and invasion ability in progesterone group were decreased.The cell proliferation,migration and invasion ability in AS group were increased.The cell proliferation,migration and invasion ability in AS+progesterone group were decreased.Compared with progesterone group,the cell proliferation,migration and invasion ability in AS+progesterone group were increased.Compared with AS group,the cell proliferation,migration and invasion ability in AS+progesterone group were decreased.(2)The protein expression results of HO-8910 cells in each group were compared.Compared with the control group,the expression of FOXO1 protein was increased,the expression ofβ-catenin protein was decreased in progesterone group.The expression of FOXO1 protein was decreased and the expression ofβ-catenin protein was increased in AS group.The expression of FOXO1 protein was decreased and the expression ofβ-catenin protein was decreased in AS+progesterone group.Compared with progesterone group,the expression of FOXO1 protein was decreased and the expression ofβ-catenin protein was increased in AS+progesterone group.Compared with AS group,the expression of FOXO1 protein had no significant difference and the expression ofβ-catenin protein was decreased in AS+progesterone group.Conclusion Progesterone could inhibit the proliferation,migration and invasion of ovarian cancer HO-8910 cells,which might be realized by promoting the protein expression of FOXO1 and inhibiting the protein expression ofβ-catenin.