目的:探讨芦荟苷抑制胃癌细胞增殖和迁移的分子机制。
方法:检测用100、200和300μg/mL芦荟素处理的人胃癌MGC-803细胞的细胞活力变化,使用CCK-8,EdU和Transwell测定的增殖和迁移能力。RT-qPCR检测细胞中HMGB1mRNA水平,HMGB1,cyclinB1,cyclinE1,E-cadherin,使用蛋白质印迹法测定MMP-2、MMP-9和p-STAT3。JASPAR数据库用于预测STAT3与HMGB1启动子的结合。在带有皮下MGC-803细胞异种移植物的BALB/c-Nu小鼠模型中,观察腹腔注射芦荟苷(50mg/kg)对肿瘤生长的影响。HMGB1,cyclinB1,cyclinE1,E-cadherin,用蛋白质印迹法检测肿瘤组织中的MMP-2、MMP-9和p-STAT3,HE染色检测肝、肺组织肿瘤转移。
结果:芦荟苷浓度依赖性地抑制MGC-803细胞的活力(P<0.05),显着减少了EdU阳性细胞数(P<0.01),细胞迁移能力减弱(P<0.01)。芦荟素剂量依赖性下调HMGB1mRNA表达(P<0.01),下调MGC-803细胞中HMGB1、细胞周期蛋白B1、细胞周期蛋白E1、MMP-2、MMP-9和p-STAT3的蛋白表达,上调E-cadherin的表达。基于JASPAR数据库的预测表明,STAT3可以与HMGB1的启动子区结合。在荷瘤小鼠中,芦荟素治疗可显著降低肿瘤体积和重量(P<0.01),降低了cyclinB1,cyclinE1,MMP-2,MMP-9,HMGB1和p-STAT3的蛋白表达,并增加了E-cadherin在肿瘤组织中的表达(P<0.01)。
结论:芦荟苷通过抑制STAT3/HMGB1信号通路减弱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 300 μg/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, E-cadherin, 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 concentration-dependently 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 up-regulated 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).
CONCLUSIONS: Aloin attenuates the proliferation and migration of gastric cancer cells by inhibiting the STAT3/HMGB1 signaling pathway.