姜黄素通过调控Notch1通路对人子宫内膜癌ISH细胞株孕激素治疗敏感性的影响

Effect of curcumin on the sensitivity of human endometrial carcinoma ISH cell line to progesterone therapy by regulating Notch1 pathway

目的探讨姜黄素是否能通过调控Notch1信号通路逆转人子宫内膜癌ISH细胞株孕激素治疗耐药性。方法醋酸甲羟孕酮(MPA)浓度递增及长期持续作用,建立人子宫内膜癌孕激素耐药细胞株ISH/MPA,MTS法检测细胞耐药指数,绘制ISH/MPA细胞生长曲线,计算细胞群体倍增时间。不同浓度姜黄素(0、7.5、15、30、60μmol/L)处理ISH/MPA细胞,MTT法检测细胞存活率。ISH/MPA细胞分为ISH/MPA组、姜黄素组、激动剂组、激动剂+姜黄素组,姜黄素组加入30μmol/L姜黄素、激动剂组加入2μmol/L NSC 22423、激动剂+姜黄素组加入30μmol/L姜黄素和2μmol/L NSC 22423,ISH/MPA组细胞不做处理,培养48 h后,流式细胞仪检测细胞凋亡率,蛋白印迹法检测Notch1、Jagged1、Hes-1蛋白表达。结果ISH/MPA细胞耐药指数随MPA浓度增加而升高,且MPA浓度为10μmol/L时,ISH和ISH/MPA细胞群体倍增时间无统计学差异,两者生长曲线基本一致。不同浓度姜黄素均降低ISH/MPA细胞存活率,且随姜黄素浓度增加而升高(P<0.05)。与ISH/MPA组比较,姜黄素组细胞凋亡率升高,Notch1、Jagged1和Hes-1蛋白表达降低,激动剂组细胞凋亡率降低,Notch1、Jagged1和Hes-1蛋白表达升高(P<0.05);与姜黄素组比较,激动剂+姜黄素组细胞凋亡率降低,Notch1、Jagged1和Hes-1蛋白表达升高(P<0.05);与激动剂组比较,激动剂+姜黄素组细胞凋亡率升高,Notch1、Jagged1和Hes-1蛋白表达降低(P<0.05)。结论姜黄素提高ISH细胞对MPA的敏感性,其可能是通过抑制Notch1/Jagged1/Hes-1信号通路发挥作用。

Objective To investigate whether curcumin can reverse the progesterone resistance of human endometrial carcinoma ISH cell lines by regulating the Notch1 signaling pathway.Methods The human endometrial carcinoma progesterone resistant cell line ISH/MPA was established by increasing the concentration and long-term persistence of medroxyprogesterone acetate(MPA).The cell resistance index was detected by MTS method,the growth curve of ISH/MPA cells was plotted and the cell population doubling time was calculated.ISH/MPA cells were treated with different concentrations of curcumin(0,7.5,15,30,60μmol/L),and the survival rate of cells was detected by MTT assay.ISH/MPA Cells were divided into ISH/MPA group,curcumin group,agonist group,agonist+curcumin group,curcumin group added 30μmol/L curcumin,agonist group added 2μmol/L NSC 22423,agonist+curcumin group added 30μmol/L curcumin and 2μmol/L NSC22423,Cells in the ISH/MPA group were not treated.After 48 h,flow cytometry instrument detected cells apoptosis rate,western blot detected Notch1,Jagged1 and Hes-1 protein expression.Results The drug resistance index of ISH/MPA cells increased with the increasing concentration of MPA,and when the concentration of MPA was 10μmol/L,the population doubling time of ISH and ISH/MPA cells had no statistical difference,and the growth curves of the two cells were basically the same.Different concentrations of curcumin decreased the survival rate of ISH/MPA cells,and increased with the increase of curcumin concentration(P<0.05).Compared with ISH/MPA group,the apoptosis rate of cells in curcumin group was increased,and the protein expressions of Notch1,Jagged1 and Hes-1 were decreased,the apoptosis rate of cells in agonist group was decreased,and the protein expressions of Notch1,Jagged1 and Hes-1 were increased(P<0.05).Compared with curcumin group,the apoptosis rate of agonist+curcumin group was decreased,and the protein expressions of Notch1,Jagged1 and Hes-1 were increased(P<0.05).Compared with agonist group,the apoptosis rate of agonist+curcumin group increased,and the protein expressions of Notch1,Jagged1 and Hes-1 decreased(P<0.05).Conclusion Curcumin enhances the sensitivity of ISH cells to MPA,possibly by inhibiting the Notch1/Jagged1/Hes-1 signaling pathway.