高糖饮食通过激活RAGE/mTOR信号通路抑制铁自噬促进结直肠癌侵袭转移  

High glucose consumption promoted invasion and migration in colorectal cancer through suppressing ferritinophagy via activating the RAGE/mTOR pathway

作　　者：熊枭 陈豪 李元亮 唐研 张好刚[1] 乔鹏飞[1] XIONG Xiao;CHEN Hao;LI Yuanliang;TANG Yan;ZHANG Haogang;QIAO Pengfei(Department of General Surgery,The Second Affiliated Hospital of Harbin Medical University,Harbin 150001,P.R.China)

机构地区：[1]哈尔滨医科大学附属第二医院普通外科,哈尔滨150001

出　　处：《中国普外基础与临床杂志》2025年第4期493-504,共12页Chinese Journal of Bases and Clinics In General Surgery

基　　金：中国光华科研基金(项目编号:YX20210615#0054)。

摘　　要：目的探讨高糖饮食环境下哺乳动物雷帕霉素靶蛋白(mechanistic target of rapamycin kinase,mTOR)/晚期糖基化终末产物受体(receptor of advanced glycation end products,RAGE)信号通路介导的铁自噬对结直肠癌(colorectal cancer,CRC)侵袭转移的影响及作用机制。方法(1)临床病例资料分析。回顾性收集2022年10月至2023年10月期间于哈尔滨医科大学附属第二医院接受手术治疗的66例CRC患者的临床资料和饮食习惯,比较高糖饮食组和正常饮食组患者的临床病理特征,并采用实时定量逆转录聚合酶链反应(real-time quantitative reverse transcription PCR,RT-qPCR)方法检测核受体辅助激活因子4(nuclear receptor coactivator 4,NCOA4)和铁蛋白(ferritin)的表达水平。(2)细胞实验。取活力良好的对数生长期HT29和HCT116结肠癌细胞,分别以含0、35、70、105、140 mmol/L葡萄糖的RPMI1640培养基培养,采用细胞计数试剂盒和乳酸脱氢酶(lactate dehydrogenase,LDH)细胞毒性检测试剂盒检测细胞生存率以及LDH活性以确定适宜浓度为105 mmol/L。再取活力良好的对数生长期HT29和HCT116细胞分组:对照组、高糖组;对照组、高糖组、靶向RAGE的siRNA转染组(si-RAGE组)、靶向RAGE的siRNA转染同时以高糖培养组(高糖+si-RAGE组);对照组、高糖组、铁自噬激活剂处理组(雷帕霉素组)、铁自噬激活剂处理同时以高糖培养组(高糖+雷帕霉素组)。对照组为未经任何处理的HT29和HCT116细胞,高糖培养组以含105 mmol/L葡萄糖的RPMI1640培养基培养细胞48 h,si-RAGE组以RAGE siRNAs转染细胞6 h,雷帕霉素组以10 nmol/L雷帕霉素处理细胞48 h,高糖+si-RAGE组以含105 mmol/L葡萄糖的RPMI1640培养基培养细胞48 h的同时以RAGE siRNAs转染细胞6 h,高糖+雷帕霉素组以含105 mmol/L葡萄糖的RPMI1640培养基培养细胞48 h的同时以10 nmol/L雷帕霉素处理细胞48 h。采用透射电镜观察铁自噬小体形成情况,采用蛋白质免疫印迹法检测RAGObjective To explore the influence and mechanism of mechanistic target of rapamycin kinase(mTOR)/receptor of advanced glycation end products(RAGE)pathway mediated-ferritinophagy on high glucose consumption promoting invasion and migration of colorectal cancer(CRC).Methods①Patients and tissue samples.Clinical data and tissues were collected from CRC patients underwent surgery and completed the dietary questionnaire in the Second Affiliated Hospital of Harbin Medical University from October 2022 to October 2023.Real-time quantitative reverse transcription PCR(qRT-PCR)was used to analyzed the expression of nuclear receptor coactivator 4(NCOA4)and ferritin in CRC and para-carcinoma tissues.②Cell culture and treatment.The HT29 and HCT116 cells were treated by RPMI1640 medium containing 0,35,70,105,140 mmol/L glucose,and cell counting kit-8(CCK-8)and lactate dehydrogenase(LDH)activity analysis were performed to confirm 105 mmol/L glucose was the optimal concentration in the current study.Then the HT29 and HCT116 cells were randomly divided into:control group,glucose group;control group,glucose group,si-RAGE group,and glucose+si-RAGE group;control group,glucose group,rapamycin group,and glucose+rapamycin group.Untreated HT29 and HCT116 cells were considered as control group.The cells in glucose group were treated with 105 mmol/L glucose for 48 h.The CRC cells in the si-RAGE group were transfected with si-RAGE for 6 h.The CRC cells in the rapamycin group were treated with 10 nmol/L rapamycin for 48 h.The CRC cells in the glucose+si-RAGE group were treated with 105 mmol/L glucose for 48 h combination transfected with si-RAGE for 6 h.The CRC cells in the glucose+rapamycin group were treated with 105 mmol/L glucose for 48 h combination treated with 10 nmol/L rapamycin for 48 h.Then electron microscopy and Western blot,wound healing assay and transwell assay were exhibited,respectively.③Azoxymethane(AOM)-induced CRC rat model.The effects of glucose consumption on malignant behavior and ferritinophagy mediated by mTOR/R

关 键 词：结直肠癌 高糖饮食 铁自噬 糖基化终末产物受体 哺乳动物雷帕霉素靶蛋白信号 

分 类 号：R735.3[医药卫生—肿瘤]