中国医科大学学报2024，Vol.53Issue(3)：207-212,6.DOI:10.12007/j.issn.0258-4646.2024.03.003
二氢青蒿素结合SERCA2b诱导结肠癌HCT-116细胞凋亡的机制研究
Mechanism of dihydroartemisinin binding SERCA2b and inducing apoptosis in colorectal cell HCT-116
摘要
Abstract
Objective To investigate the binding sites of dihydroartemisinin(DHA)and sarcoplasmic/endoplasmic reticulum calcium ATPase(SERCA)and explore the mechanisms underlying apoptosis induction in HCT-116 colon cancer cells through the mitochondrial pathway.Methods HCT-116 cells were cultured in DHA concentrations of 0,10,20,40 μmol/L.After 24 h of culture,Western blot-ting assessed SERCA concentration,CCK-8 measured cell proliferation,Hoechst nuclear staining examined cell apoptosis,JC-1 probe evaluated mitochondrial membrane potential.LeDock molecular docking predicted DHA and SERCA binding sites.Synthetic SERCA2b mutated and non-mutated proteins at Ile315 and Thr316 sites were combined with small molecule DHA using biofilm interference tech-nology.Results Western blotting revealed a significant decrease in SERCA2 protein levels with increasing DHA concentration.CCK-8 demonstrated a statistically significant decrease in cell proliferation with increasing DHA concentration(P＜0.01).Hoechst nuclear staining illustrated DHA-induced rounding and pyknosis of HCT-116 cell nuclei compared to the control group.DHA gradually decreased mitochondrial membrane potential within the first 4 h of treatment,starting from 5 h,followed by a sustained reduction.Molecular docking predicted a hydrogen bond with Thr316 and a hydrophobic interaction with Ile315.Biofilm interference techniques indicated robust binding of DHA to non-mutated SERCA2b protein,while binding to the SERCA2b mutant protein at Ile315 and Thr316 sites was poor.Conclusion DHA directly binds to the Ile315 and Thr316 sites of SERCA2b,inducing apoptosis in colon cancer cells HCT-116 through the mitochondrial pathway.关键词
二氢青蒿素/肌浆/内质网钙ATP酶/结直肠肿瘤Key words
dihydroartemisinin/sarcoplasmic/endoplasmic reticulum calcium ATPase/colorectal cancer分类
医药卫生引用本文复制引用
孙卢浩然,卢敏..二氢青蒿素结合SERCA2b诱导结肠癌HCT-116细胞凋亡的机制研究[J].中国医科大学学报,2024,53(3):207-212,6.基金项目
辽宁省科技厅2023年支持中国医科大学高质量发展资金项目计划(辽科发规[2023]61号) （辽科发规[2023]61号）
