摘要
目的:探究七叶皂苷对氧化三甲胺(TMAO)诱导的结直肠癌细胞株增殖及迁移侵袭的影响,并探讨其作用机制.方法:取结直肠癌HCT116细胞系,分为对照组、TMAO组、观察组3组,并分别进行处理.对照组不做处理,TMAO组仅使用100 µmol/L TMAO诱导,观察组使用同剂量TMAO诱导后再进行10 μmol/L七叶皂苷治疗处理.随后使用四甲基偶氮唑盐比色(MTT)法、划痕愈合实验及细胞迁移侵袭试验技术(Transwell)实验分别检测细胞的增殖、迁移及侵袭行为,并使用蛋白质印迹(Western Blotting)实验检测SRC、pBTK、pTRIO蛋白表达情况.结果:TMAO诱导可增强HCT116增殖及迁移侵袭能力,并显著上调SRC-BTK-TRIO通路相关基因的mRNA水平,而七叶皂苷干预能抑制癌细胞增殖(P＜0.01)并降低其迁移(P＜0.001)、侵袭(P＜0.01)能力,同时下调 SRC(P＜0.05)、BTK(P＜0.01)、TRIO(P＜0.05)蛋白的表达.结论:七叶皂苷可通过下调SRC-BTK-TRIO通路显著抑制HCT116细胞增殖及迁移侵袭能力.
Abstract
Objective:To investigate the effect of escin on the proliferation,migration,and invasion of colorectal cancer cell line HCT116 induced by tri methylamine N-oxide(TMAO)and to explore its mechanism of action.Methods:HCT116 colorectal cancer cells were divided into a control group,a TMAO group,and an experimental group.The control group received no treatment,the TMAO group was induced with 100 µmol/L TMAO,and the experimental group was treated with 100 μmol/L TMAO followed by 10 µmol/L escin.Cell proliferation,migration,and invasion were assessed using the MTT assay,scratch wound healing assay,and Transwell migration and invasion assays,respectively.The expression levels of SRC,pBTK,and pTRIO proteins were measured by Western blotting.Results:TMAO induction enhanced the proliferation,migration,and invasion of HCT116 cells and significantly upregulated the mRNA levels of genes in the SRC-BTK-TRIO pathway.Escin treatment inhibited cancer cell proliferation(P＜0.01),reduced their migration(P＜0.001)and invasion(P＜0.01)capabilities,and downregulated the expression of SRC(P＜0.05),BTK(P＜0.01),and TRIO(P＜0.05)proteins.Conclusion:Escin can significantly inhibit the proliferation,migration,and invasion of HCT116 cells by downregulating the SRC-BTK-TRIO pathway.
基金项目
国家自然科学基金面上项目(81871816)
湖北省卫生健康委员会面上项目(WJ2021M157)
湖北省自然科学基金项目(2020CFB661)
武汉市卫生健康委员会面上项目(WX20B14)
武汉大学医学部教学研究项目(2020028)
维普
万方数据