文献类型：期刊文献

中文题名：姜黄素调节LKB1-AMPK-LC3信号通路对膀胱癌细胞增殖、迁移和顺铂耐药性的影响

英文题名：Impacts of curcumin on proliferation,migration and cisplatin resistance of bladder cancer cells by regulating LKB1-AMPK-LC3 signaling pathway

作者：王乾[1];赵黎明[2]

机构：[1]上海健康医学院附属崇明医院,上海202150;[2]华东理工大学生物工程学院,生物反应器国家重点实验室,上海200237

年份：2025

卷号：41

期号：1

起止页码：9

中文期刊名：细胞与分子免疫学杂志

外文期刊名：Chinese Journal of Cellular and Molecular Immunology

收录：;北大核心:【北大核心2023】；

基金：上海市崇明区“可持续发展科技创新行动计划”项目(CKY2021-14)。

语种：中文

中文关键词：姜黄素;LKB1-AMPK-LC3;膀胱癌;增殖;迁移;顺铂耐药性

外文关键词：Curcumin;LKB1-AMPK-LC3;bladder cancer;proliferation;migration;cisplatin resistance

摘要：目的研究姜黄素调节肝激酶B1-磷酸腺苷激活的蛋白激酶-微管相关蛋白1轻链3(LKB1-AMPK-LC3)信号通路对膀胱癌细胞增殖、迁移和顺铂(DDP)耐药性的影响。方法体外培养人膀胱癌细胞系T24细胞,用DDP诱导建立其DDP耐药细胞T24/DDP细胞,用不同浓度姜黄素处理T24和T24/DDP细胞后以MTT法筛选姜黄素最佳作用浓度。将T24细胞随机分为对照组、姜黄素组、二甲双胍组、姜黄素联合二甲双胍组,用姜黄素和LKB1-AMPK激活剂二甲双胍分组处理后以MTT法、单丹磺酰戊二胺(MDC)荧光染色法、细胞划痕法、流式细胞术分别检测各组T24细胞增殖、自噬、迁移与凋亡;Western blot法检测各组T24细胞LKB1-AMPK-LC3信号通路相关蛋白表达。将T24/DDP细胞随机分为对照组、姜黄素组、二甲双胍组、姜黄素联合二甲双胍组,用姜黄素和二甲双胍分组处理的同时用不同浓度DDP处理,然后以MTT法检测姜黄素对T24/DDP细胞DDP耐药系数的影响。将T24/DDP细胞随机分为对照组、DDP组、DDP联合姜黄素组、DDP联合二甲双胍组、DDP和姜黄素联合二甲双胍组,用DDP、姜黄素和二甲双胍分组处理后以同样方法检测各组T24/DDP细胞增殖、自噬、迁移、凋亡、耐药与LKB1-AMPK-LC3信号通路相关蛋白表达。结果相比对照组,姜黄素组T24细胞活力、自噬体相对数、迁移率、磷酸化的LKB1(p-LKB1)/LKB1、磷酸化的AMPK(p-AMPK)/AMPK与LC3Ⅱ/LC3Ⅰ、T24/DDP细胞DDP耐药系数降低,T24细胞凋亡率升高;二甲双胍组各指标变化与姜黄素组相反。相比姜黄素组,姜黄素联合二甲双胍组T24细胞活力、自噬体相对数、迁移率、p-LKB1/LKB1、p-AMPK/AMPK与LC3Ⅱ/LC3Ⅰ、T24/DDP细胞DDP耐药系数升高,T24细胞凋亡率降低。相比对照组,DDP组T24/DDP细胞各指标无明显变化。相比对照组、DDP组,DDP联合姜黄素组T24/DDP细胞活力、自噬体相对数、迁移率、P-糖蛋白(P-gp)蛋白表达、p-LKB1/LKB1、p-AMPK/AMPK与LC3Ⅱ/LC3Ⅰ降低,T24/DDP细胞凋亡率升高;DDP联合二甲双胍组上述各指标变化与DDP联合姜黄素组相反。相比DDP联合姜黄素组,DDP和姜黄素联合二甲双胍组T24/DDP细胞活力、自噬体相对数、迁移率、P-gp蛋白表达、p-LKB1/LKB1、p-AMPK/AMPK与LC3Ⅱ/LC3Ⅰ升高,T24/DDP细胞凋亡率降低。结论姜黄素可降低LKB1-AMPK-LC3信号通路活性,进而抑制膀胱癌细胞自噬、增殖与迁移,促使其凋亡,并减弱其DDP耐药性。
Objective To study the impacts of curcumin on the proliferation,migration and cisplatin(DDP)resistance of bladder cancer cells by regulating the liver kinase B1-AMP activated protein kinase-microtubule-associated protein 1 light chain 3(LKB1-AMPK-LC3)signaling pathway.Methods Human bladder cancer cell line T24 was cultured in vitro,and its DDP resistant T24/DDP cells were induced by cisplatin(DDP).After treating T24 and T24/DDP cells with different concentrations of curcumin,the optimal concentration of curcumin was screened by MTT assay.T24 cells were randomly grouped into control group,curcumin group,metformin group,and combination group of curcumin and metformin.After treatment with curcumin and LKB1-AMPK activator metformin,the proliferation,autophagy,migration,and apoptosis of T24 cells in each group were detected by MTT assay,monodansylcadavrine(MDC)fluorescence staining,cell scratch assay,and flow cytometry,respectively.Western blot was used to detect the expression of proteins related to LKB1-AMPK-LC3 signaling pathway in T24 cells of each group.T24/DDP cells were randomly assigned into control group,curcumin group,metformin group,and combination group of curcumin and metformin.Cells were treated with curcumin and metformin according to grouping and treated with different concentrations of DDP simultaneously.Then,the effect of curcumin on the DDP resistance coefficient of T24/DDP cells was detected by MTT assay.T24/DDP cells were randomly grouped into control group,DDP group,combination groups of DDP and curcumin,DDP and metformin,DDP,curcumin and metformi.After treatment with DDP,curcumin,and metformin,the proliferation,autophagy,migration,apoptosis,drug resistance,and the expression of proteins related to LKB1-AMPK-LC3 signaling pathway in T24/DDP cells of each group were detected with the same methods.Results Compared with the control group,the activity of T24 cells,relative number of autophagosomes,migration rate,Phosphorylated-LKB1(p-LKB1)/LKB1,Phosphorylated-AMPK(p-AMPK)/AMPK,LC3Ⅱ/LC3Ⅰ,and the DDP resistance coefficient of T24/DDP cells in the curcumin group were lower,and the apoptosis rate of T24 cells was higher;the changes in various indicators in the metformin group were opposite to those in the curcumin group.Compared with the curcumin group,the activity of T24 cells,relative number of autophagosomes,migration rate,p-LKB1/LKB1,p-AMPK/AMPK,LC3II/LC3I,and the DDP resistance coefficient of T24/DDP cells in the combination group of curcumin and metformin were higher,and the apoptosis rate of T24 cells was lower.Compared with the control group,there were no obvious changes in various indicators of T24/DDP cells in the DDP group.Compared with the control group and DDP group,the viability of T24/DDP cells,relative number of autophagosomes,migration rate,P-glycoprotein(P-gp)protein expression,p-LKB1/LKB1,p-AMPK/AMPK,and LC3Ⅱ/LC3Ⅰin the combination group of DDP and curcumin were lower,and the apoptosis rate of T24/DDP cells was higher;the changes in the above indicators in the combination group of DDP and metformin were opposite to those in the combination group of DDP and curcumin.Compared with the combination group of DDP and curcumin,the viability of T24/DDP cells,relative number of autophagosomes,migration rate,P-gp protein expression,p-LKB1/LKB1,p-AMPK/AMPK,and LC3Ⅱ/LC3Ⅰin the combination group of DDP,curcumin and metformin were higher,and the apoptosis rate of T24/DDP cells was lower.Conclusion Curcumin can reduce the activity of LKB1-AMPK-LC3 signaling pathway,thereby inhibiting autophagy,proliferation and migration of bladder cancer cells,promoting their apoptosis,and weakening their resistance to DDP.