自噬及AKT在姜黄素增强卵巢癌细胞卡铂敏感性的机制研究

Mechanism of autophagy and AKT in curcumin enhancing carboplatin sensitivity in ovarian cancer cells

目的本实验拟研究蛋白激酶B(AKT)、自噬相关基因Beclin1、微管相关蛋白1轻链3(LC3)及肿瘤坏死因子受体相关蛋白1(TRAP1)在姜黄素逆转卡铂耐药中的机制。方法培养卵巢癌转基因细胞T2,使用质粒法敲减AKT。采用CCK-8法检测细胞增殖,Western blot检测姜黄素及卡铂作用前后各指标的变化。结果姜黄素及卡铂抑制T2细胞增殖,降低AKT蛋白表达,两药联用时增加LC3、TRAP1蛋白表达。敲减AKT后Beclin1、LC3、TRAP1的表达水平增加,细胞对卡铂的敏感性提高。姜黄素与卡铂对T2细胞有协同抑制作用,敲减AKT后协同作用消失。结论姜黄素可以提高T2细胞的卡铂敏感性,与AKT通路及自噬水平增加有关。AKT是姜黄素逆转T2细胞卡铂耐药的关键因素。

Objective To investigate the mechanism of AKT protein and Beclin1, LC3 and TRAP1 in the reversal of carboplatin resistance by curcumin. Methods The ovarian cancer transgenic cell line T2 was cultured, and the AKT protein level of T2 cells was knocked down by plasmid method. The proliferation activity of the cells was detected by CCK-8 method, and the changes of each index were detected by Western blot. Results Curcumin and carboplatin inhibit the proliferation of T2 cells and reduce the expression of AKT. The combination of the two drugs increased the expression of LC3 and TRAP1. After knocking down AKT, the expression levels of Beclin1, LC3 and TRAP1 increased, and the sensitivity of cells to carboplatin increased. Curcumin and carboplatin have a synergistic inhibitory effect on T2 cells, and the synergistic effect disappears after knocking down AKT. Conclusion Curcumin improves the carboplatin sensitivity of T2 cells and is related to the increase of AKT pathway and autophagy. AKT is a key factor for curcumin to reverse the carboplatin resistance of T2 cells.