摘要
目的探究二甲双胍(Met)对胃癌细胞增殖、迁移、侵袭的调控机制。方法胃癌细胞株SGC7901和BGC823经培养并进行Met干预,CCK-8检测细胞的增殖能力,Tanswell实验检测细胞迁移和侵袭能力,RT-PCR及Westernblot检测上皮间质转化(EMT)标志物E-cadherin和Vimentin的表达,以及RAGE/NOXs信号通路的表达。慢病毒转染构建过表达RAGE细胞系SGC7901-RAGE细胞,建立裸鼠胃癌模型,并行Met治疗,肿瘤体积每3天测量1次,28 d后,肿瘤组织称重,并进行免疫组化染色和Western blot检测,验证Ki-67,以及RAGE、NOXs的表达水平。结果体外实验发现,Met能有效抑制胃癌细胞的增殖、迁移和侵袭能力。同时,Met增加E-cadherin的mRNA和蛋白表达,降低Vimentin的mRNA和蛋白表达。Met可以限制RAGE、NOX4的表达。体内实验证实腹腔注射Met能抑制过表达RAGE诱导的胃癌细胞过度增长,免疫组化结果显示Met能降低Ki-67阳性率。Western blot显示,Met可以抑制RAGE和NOX4在肿瘤组织中的表达。结论Met可以调控RAGE/NOXs信号通路逆转EMT抑制胃癌侵袭转移。
Objective To explore the regulatory mechanism of metformin(Met)on the proliferation,migration,and invasion of gastric cancer.Methods Gastric cancer cell lines SGC7901 and BGC823 were cultured and subjected to Met intervention.Cell count kit-8(CCK-8)was used to detect cell proliferation ability,tanswell assay was used to detect the cell migration and invasion ability;RT-PCR and Western blot were uesd to detect the expressions of epit-helial-mesenchymal transition(EMT)markers E-cadherin and Vimentin,and the expression of the RAGE/NOXs sig-naling pathway.Construction of overexpressing RAGE cell line SGC7901-RAGE by lentivirus transfection,and es-tablish the nude mouse gastric cancer model,treat with Met.Tumor volume was measured every three days,and weighed after 28 days;immunohistochemical and western blot were uesd to detect the expressions of Ki-67,RAGE and NOX4.Results In vitro experiments,Met treatment could effectively inhibit the proliferation,migration,and invasion abilities of SGC7901 and BGC823.Meanwhile,Met could increase the mRNA and protein expression of E-cadherin,and reduce the mRNA and protein expression of Vimentin.Met could suppress the expressions of RAGE and NOX4.In vivo experiments,intraperitoneal injection of Met could inhibit the excessive growth of gastric cancer cells induced by RAGE overexpression.Immunohistochemical showed that Met could reduce the positive rate of Ki-67.Western blot showed Met could inhibit the expressions of RAGE and NOX4 in tumor tissue.Conclusions Met inhibited gastric cancer migration,invasion and reverse EMT by regulating the RAGE/NOXs signaling pathway.
作者
金丹
郭群依
金剑英
谢静静
JIN Dan;GUO Yiqun;JIN Jianying;XIE Jingjing(Taizhou Hospital of Zhejiang Province,Taizhou 3I7000,Zhejang,China)
出处
《现代实用医学》
2025年第2期120-124,F0003,共6页
Modern Practical Medicine
基金
台州市科技计划项目(21ywa19)。
