桦木酸对胃癌MGC-803细胞迁移和侵袭的抑制作用及其机制  

Inhibitory effects of betulinic acid on migration and invasion of gastric cancer MGC-803 cells and their mechanisms

作　　者：许广松 蒋海兵 盘箐[3] 李国庆 XU Guangsong;JIANG Haibing;PAN Jing;LI Guoqing(Department of General surgery,Second Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001,China;Department of Gastrointestinal Medicine,Second Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001,China;Department of Pathogenic Biology and Immunology,Yongzhou Vocational and Technical College,Yongzhou 425100,China)

机构地区：[1]南华大学衡阳医学院附属第二医院普外科,湖南衡阳421001 [2]南华大学衡阳医学院附属第二医院消化内科,湖南衡阳421001 [3]永州职业技术学院病原生物与免疫学教研室,湖南永州425100

出　　处：《吉林大学学报（医学版）》2022年第1期122-128,共7页Journal of Jilin University：Medicine Edition

基　　金：湖南省卫计委2017年度第二批科研计划项目(A2017014);湖南省卫健委科研项目(202103032022);湖南省衡阳市科技局指导性项目(139)。

摘　　要：目的:探讨桦木酸(BEA)对胃癌MGC-803细胞增殖、凋亡、迁移和侵袭的影响,并阐明其作用机制。方法:MGC-803细胞分为对照组和不同剂量BEA组,分别采用含0、2.5、5.0、10.0、20.0、40.0和80.0μmol·L^(-1) BEA的DMEM高糖培养基常规培养。采用CCK-8法、流式细胞术、划痕实验和Transwell法分别检测MGC-803细胞增殖率、凋亡率、迁移率和侵袭细胞数;Westernblotting法检测各组MGC-803细胞中SMAD同源物7(SMAD7)、转化生长因子β受体1(TGF-βR1)、磷酸化SMAD同源物2(p-SMAD2)、磷酸化SMAD同源物3(p-SMAD3)、性别决定区Y框蛋白4(SOX4)、E盒结合锌指蛋白2(ZEB2)、基质金属蛋白酶9(MMP-9)、Snail和Slug蛋白表达水平。结果:分别培养24、48和72 h后,与对照组比较,2.5、5.0、10.0、20.0、40.0和80.0μmol·L^(-1) BEA组细胞增殖率明显降低(P<0.05);培养72 h后,与对照组比较,2.5、5.0、10.0和20.0μmol·L^(-1) BEA组细胞凋亡率明显升高(P<0.05);培养24 h后,与对照组比较,2.5、5.0、10.0和20.0μmol·L^(-1) BEA组细胞迁移率明显降低(P<0.05),侵袭细胞数明显减少(P<0.05)。与对照组比较,培养48 h后,20μmol·L^(-1) BEA组细胞中SMAD7蛋白表达水平明显升高(P<0.05),TGF-βR1、p-SMAD2、p-SMAD3、SOX4、ZEB2、MMP-9、Snail和Slug蛋白表达水平明显降低(P<0.05)。结论:BEA通过上调SMAD7表达以及抑制TGF-β/SMAD信号通路激活,调节胃癌细胞增殖、凋亡、迁移和侵袭。Objective:To investigate the effects of betulinic acid(BEA)on the proliferation,apoptosis,migration and invasion of gastric cancer MGC-803 cells,and to elucidate their mechanisms.Methods:The MGC-803 cells were divided into control group and different doses of BEA groups,and the cells were cultured with DMEH high sugar medium containing 0,2.5,5.0,10.0,20.0,40.0,and 80.0μmol·L^(-1) BEA.The proliferation rates,apoptotic rates,migration rates and the number of invasion MGC-803 cells were detected by CCK-8 method,flow cytometry,scratch test and Transwell method,respectively.The expression levels of SMAD homology 7(SMAD7),transforming growth factor-βreceptor 1(TGF-βR1),phosphorylated SMAD2(p-SMAD2),phosphorylated SMAD3(p-SMAD3),SRY-box transcription factor 4(SOX4),zinc finger E-box-binding protein 2(ZEB2),matrix metalloproteinase-9(MMP-9),Snail and Slug proteins in the MGC-803 cells in various groups were detected by Western blotting method.Results:Compared with control group,the proliferation rates of cells in 2.5,5.0,10.0,20.0,40.0 and 80.0μmol·L^(-1) BEA groups were decreased after 24,48 and 72 h of incubation(P<0.05).After 72 h of incubation,the apoptotic rates in 2.5,5.0,10.0 and 20.0μmol·L^(-1) BEA groups were increased compared with control group(P<0.05).After 24 h of incubation,compared with control group,the migration rates in 2.5,5.0,10.0 and 20.0μmol·L^(-1) BEA groups were decreased(P<0.05),and the number of invasion cells was decreased(P<0.05).The results of Western blotting method revealed that compared with control group,the expression level of SMAD7 protein in 20μmol·L^(-1) BEA group was increased(P<0.05),and the expression levels of TGF-βRI,p-SMAD2,p-SMAD3,SOX4,ZEB2,MMP-9,Snail and Slug proteins were decreased after 48 h of incubation.Conclusion:BEA regulates the proliferation,apoptosis,migration and invasion of gastric cancer cells by up-regulating the expression of SMAD7 and inhibiting the activation of TGF-β/Smad signaling pathway.

关 键 词：桦木酸 胃肿瘤 流式细胞术 细胞增殖 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]