芦荟苷可抑制胃癌细胞的增殖和迁移:基于下调STAT3/HMGB1信号通路  

Aloin inhibits gastric cancer cell proliferation and migration by suppressing the STAT3/HMGB1 signaling pathway

作　　者：葛菲 万梦琪 程振宇 陈雪雷 陈芊伊 戚之琳 GE Fei;WAN Mengqi;CHENG Zhenyu;CHEN Xuelei;CHEN Qianyi;QI Zhilin(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Wannan Medical College,Wuhu 241002,China;Anhui Provincial Key Laboratory of Active Biological Macromolecules,Wannan Medical College,Wuhu 241002,China;School of Pharmacy,Wannan Medical College,Wuhu 241002,China;School of Clinical Medicine,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院基础医学院生物化学与分子生物学教研室,安徽芜湖241002 [2]皖南医学院活性生物大分子安徽省重点实验室,安徽芜湖241002 [3]皖南医学院药学院,安徽芜湖241002 [4]皖南医学院临床学院,安徽芜湖241002

出　　处：《南方医科大学学报》2023年第5期702-709,共8页Journal of Southern Medical University

基　　金：安徽高校自然科学研究项目重大项目(KJ2020ZD54);2021年学科(专业)拔尖人才学术资助项目(gxbjZD2021060);活性生物大分子研究安徽省重点实验室项目(1306C083008);安徽省大学生创新创业训练计划项目(S202110368109);2021年度校大学生科研资助金项目(WK2021XS07,WK2021XS64)。

摘　　要：目的通过体内外实验探讨芦荟苷抑制胃癌细胞增殖和迁移的分子机制。方法胃癌MGC-803细胞分为对照组、不同浓度芦荟苷(100、200和300μg/mL)处理组。CCK-8、EdU和Transwell实验分别检测细胞活力、细胞增殖和迁移能力;RT-qPCR检测HMGB1 mRNA的表达水平;Western blot检测HMGB1,Cyclin B1/E1,E-cadherin,MMP-2/9的表达以及STAT3的磷酸化。JASPAR数据库预测STAT3与HMGB1启动子的结合。MGC-803细胞(2×106/只)注射于BALB/c-Nu小鼠腋下进行皮下植瘤,裸鼠随机分为对照组和芦荟苷组(n=5)。实验组使用芦荟苷50 mg/kg/d腹腔注射,对照组注射等量PBS。每日测量瘤体大小和小鼠质量;提取肿瘤组织蛋白,Western blot检测HMGB1,Cyclin B1/E1,E-cadherin,MMP-2/9和p-STAT3的表达。HE染色检测肝、肺组织中肿瘤转移情况。结果芦荟苷能够浓度依赖性的抑制胃癌细胞活力(P<0.05),不同浓度芦荟苷处理组EdU阳性染色细胞与对照组相比显著减少(P<0.01),芦荟苷处理的胃癌细胞转移能力明显较对照组减弱(P<0.01)。芦荟苷能够浓度依赖性的下调HMGB1 mRNA和蛋白表达(P<0.01),下调Cyclin B1,E1,MMP-2/9,上调E-cadherin,明显抑制p-STAT3。JASPAR数据库预测STAT3能够与HMGB1启动子区域相结合。芦荟苷处理组瘤体大小和质量明显低于对照组(P<0.01)。芦荟苷处理组Cyclin B1/E1,MMP-2/9,HMGB1和p-STAT3的表达与对照组相比明显降低,E-cadherin的表达则显著增强(P<0.01)。结论芦荟苷通过抑制p-STAT3/HMGB1信号途径,抑制胃癌细胞的增殖和迁移。Objective To investigate the molecular mechanism underlying the inhibitory effect of aloin on the proliferation and migration of gastric cancer cells.Methods Human gastric cancer MGC-803 cells treated with 100,200 and 300μg/mL aloin were examined for changes in cell viability,proliferation and migration abilities using CCK-8,EdU and Transwell assays.HMGB1 mRNA level in the cells was detected with RT-qPCR,and the protein expressions of HMGB1,cyclin B1,cyclin E1,Ecadherin,MMP-2,MMP-9 and p-STAT3 were determined using Western blotting.JASPAR database was used to predict the binding of STAT3 to HMGB1 promoter.In a BALB/c-Nu mouse model bearing subcutaneous MGC-803 cell xenograft,the effect of intraperitoneal injection of aloin(50 mg/kg)on tumor growth was observed.The protein expressions of HMGB1,cyclin B1,cyclin E1,E-cadherin,MMP-2,MMP-9 and p-STAT3 in the tumor tissue was examined using Western blotting,and tumor metastasis in the liver and lung tissues was detected using HE staining.Results Treatment with aloin concentrationdependently inhibited the viability of MGC-803 cells(P<0.05),significantly reduced the number of EdU-positive cells(P<0.01),and attenuated the migration ability of the cells(P<0.01).Aloin treatment dose-dependently down-regulated HMGB1 mRNA expression(P<0.01),lowered the protein expressions of HMGB1,cyclin B1,cyclin E1,MMP-2,MMP-9 and p-STAT3,and upregulated E-cadherin expression in MGC-803 cells.Prediction based on JASPAR database suggested that STAT3 could bind to the promoter region of HMGB1.In the tumor-bearing mice,aloin treatment significantly reduced the tumor size and weight(P<0.01),lowered the protein expressions of cyclin B1,cyclin E1,MMP-2,MMP-9,HMGB1 and p-STAT3 and increased the expression of E-cadherin in the tumor tissue(P<0.01).Conclusion Aloin attenuates the proliferation and migration of gastric cancer cells by inhibiting the STAT3/HMGB1 signaling pathway.

关 键 词：芦荟苷 高迁移族盒蛋白1 胃癌 增殖 迁移 

分 类 号：R285[医药卫生—中药学]