井冈山大学学报（自然科学版）2025，Vol.46Issue(1)：51-57,7.DOI:10.3969/j.issn.1674-8085.2025.01.008
双氢青蒿素经AMPK/mTOR通路逆转卵巢癌细胞A2780/DDP顺铂耐药的机制研究
REVERSAL EFFECT AND ITS MECHANISM OF DIHYDROARTEMISININ ON CISPLATIN RESISTANCE IN OVARIAN CANCER CELL LINES A2780/DDP BY AMPK/MTOR PATHWAY
摘要
Abstract
In order to explore the molecular mechanism of dihydroartemisinin(DHA)reversing the cisplatin resistance of ovarian cancer cells A2780/DDP through AMPK/mTOR pathway.Human ovarian cancer A2780 and cisplatin-resistant A2780/DDP cells were cultured with DHA under different concentrations(5,10,20,40,80 μmol/L)in vitro.The CCK-8 method was used to calculate the inhibitory effect of DHA on the proliferation of A2780 and A2780/DDP cells after 48 hours,and the most appropriate DHA concentration was selected for the subsequent experiments.There were single DDP groups(0.625,1.25,2.5,5 and 10 mg/L)and the groups of DHA combined with cisplatin of each concentration.The IC50 of cisplatin and the resistance reversal fold of DHA on A2780 and A2780/DDP cells were calculated.DAPI assay was used to detect the effect of DHA on apoptosis of A2780/DDP.Effect of DHA on AMPKα,mTOR,p-mTOR,HIF-1α,and P-gp in A2780/DDP cells were detected by Western blot after treated with AMPKα siRNA and the agonist of AMPK.The results showed that the growth of A2780/DDP cells was significantly inhibited with increasing DHA concentration in a concentration-dependent manner.And the optimal concentration of DHA to reverse drug resistance was 10 μmol/L.The IC50 of cisplatin against A2780/DDP in the DDP+DHA group was 4.95 mg/L and 20.33 mg/L in the DDP group.And the resistance reversal fold of DHA was 4.11.The DDP+DHA group showed apoptotic morphology with reduced size,nuclear consolidation or fragmentation,bright blue color and significantly increased apoptosis rate.After knocking down the expression of AMPK gene,the expression of mTOR phosphorylation,HIF-1α and P-gp was increased,and the growth rate of A2780/DDP cells was significantly increased.Meanwhile,after using mTOR,HIF-1α and P-gp agonists respectively,the expression of their downstream HIF-1α and P-gp increased significantly,and the growth rate of A2780/DDP cells increased significantly compared with the DHA group.DHA may inhibit the proliferation and promote the apoptosis of A2780/DDP cells by regulating AMPK/mTOR signaling pathway and down-regulating the expression of HIF-1α and P-gp to reverse the drug resistance.关键词
双氢青蒿素/卵巢癌/顺铂耐药/AMPK/mTOR信号通路Key words
DHA/ovarian cancer/cisplatin resistance/AMPK/mTOR signaling pathway分类
生物学引用本文复制引用
童焰,肖沁,赖智清,刘朝霞..双氢青蒿素经AMPK/mTOR通路逆转卵巢癌细胞A2780/DDP顺铂耐药的机制研究[J].井冈山大学学报（自然科学版）,2025,46(1):51-57,7.基金项目
国家自然科学基金项目(82260298) （82260298）
江西省中医院管理局科技计划项目(2021A228) （2021A228）
