羽扇豆醇对结直肠癌HCT116细胞的抑制作用及机制研究  

Inhibitory effect of lupoel on colorectal cancer HCT116 cells and its mechanism

作　　者：韩泽平 蔡贞[1] HAN Zeping;CAI Zhen(Department of Clinical Laboratory,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong 510515,China;Central Laboratory,Panyu Central Hospital,Guangzhou,Guangdong 511400,China)

机构地区：[1]南方医科大学南方医院检验科,广东广州510515 [2]广州市番禺区中心医院中心实验室,广东广州511400

出　　处：《国际检验医学杂志》2022年第20期2433-2440,共8页International Journal of Laboratory Medicine

基　　金：广东省中医药局科研项目(20192073);广东省广州市中医药和中西医结合科技项目(20212A010025);广东省广州市番禺区科技计划项目(2020-Z04-026)。

摘　　要：目的探讨羽扇豆醇对结直肠癌HCT116细胞的抑制作用及潜在分子机制。方法体外培养结直肠癌HCT116细胞,采用CCK-8法、克隆形成试验、划痕愈合试验、Transwell试验及流式细胞术评估不同浓度羽扇豆醇对HCT116细胞的增殖,克隆形成、迁移和侵袭能力,以及凋亡的影响;进一步通过TMT标记定量蛋白质组学分析及生物信息学分析羽扇豆醇抑制HCT116细胞生长的潜在机制,并运用蛋白质免疫印迹法验证相关蛋白的表达水平。结果羽扇豆醇具有抑制HCT116细胞增殖、克隆形成、迁移和侵袭能力,以及促进细胞凋亡,并且均呈浓度依赖性。羽扇豆醇干预HCT116细胞后共发现490种蛋白表达差异明显。经生物信息学分析发现,羽扇豆醇主要参与了HCT116细胞的分裂、增殖、凋亡及营养物质代谢等生物学过程,可能与糖酵解/糖异生、缺氧诱导因子(HIF)-1信号通路、淀粉和蔗糖代谢途径等信号通路有关。蛋白质免疫印迹法结果显示,羽扇豆醇可下调HIF-1α、α-烯醇化酶(ENO1)和乳酸脱氢酶A(LDHA)蛋白表达水平。结论羽扇豆醇对人结直肠癌HCT116细胞的生长及转移有明显抑制作用,其机制与下调糖酵解信号通路中HIF-1α、LDHA和ENO1蛋白的表达水平有关。Objective To investigate the inhibitory effect and potential molecular mechanism of lupeol on colorectal cancer HCT116 cells.Methods Colorectal cancer HCT116 cells were cultured in vitro,and the effect of different concentrations of lupinol on the proliferation,clone formation,migration and invasion,and apoptosis of HCT116 cells were evaluated by CCK-8 assay,clone formation assay,wound healing assay,Transwell assay and flow cytometry.The potential mechanism of lupeol inhibiting the growth of HCT116 cells was further analyzed by TMT quantitative proteomics analysis and bioinformatics,and the expression levels of related proteins was verified by Western blotting.Results Lupeol inhibited the proliferation,clone formation,migration and invasion of colorectal cancer HCT116 cells and promoted apoptosis of cells in a concentration-dependent manner.A total of 490 proteins were found to express differently after the treatment of lupeol on HCT116 cells.Bioinformatics analysis indicated that lupeol was mainly involved in biological processes such as division,proliferation,apoptosis and nutrient metabolism of HCT116 cells,and might be related to glycolysis/gluconeogenesis,hypoxia inducible factor(HIF)-1 signaling pathway,starch and sucrose metabolism pathway and other signaling pathways.Western blotting results showed that lupeol could down-regulate HIF-1α,α-enolase(ENO1)and lactate dehydrogenase-A(LDHA)proteins expression levels.Conclusion Lupeol has a significant inhibitory effect on the growth and metastasis of human colorectal cancer HCT116 cells,and its mechanism might be related to the down-regulation of the expression levels of HIF-1α,LDHA and ENO1 proteins in the glycolysis signaling pathway.

关 键 词：羽扇豆醇 结直肠癌HCT116细胞 TMT标记定量蛋白质组学分析 生物信息学分析 糖酵解 

分 类 号：R735.3[医药卫生—肿瘤] R446.1[医药卫生—临床医学]