DADS上调miR-7通过EGFR/Rac1/Pak1信号通路抑制胃癌SGC-7901细胞增殖、迁移侵袭与EMTOA
DADS up-regulates miR-7 to inhibit proliferation,migration,invasion and EMT of gastric cancer SGC-7901 cells through EGFR/Rac1/Pak1 signaling pathway
目的 研究DADS下调miR-7通过EGFR/Rac1/Pak1信号通路抑制SGC-7901细胞EMT与迁移侵袭及其分子机制.方法 慢病毒p HBLV-U6-RFP-T2A-Puro-hsa-miR-7-antago及对照空载体p HBLV-U6-RFP-T2A-Puro分别转染SGC-7901建立稳定沉默miR-7的SGC-7901细胞系;MTT、划痕、迁移和侵袭实验检测细胞增殖、迁移和侵袭;qRT-PCT、Western blot与免疫荧光检测EGFR、Rac1、Pak1、Vimentin、E-cadherin表达.结果 成功构建稳定沉默miR-7的SGC-7901细胞,qRT-PCR显示,转染慢病毒si-mir7的SGC7901中miR-7的表达明显下调.DADS处 理 后,SGC-7901细胞中miR-7的表达明显上调.24、48和72 h后,miR-7沉默组增殖率较SGC7901组与vector组明显增加.DADS处理后各组增殖率较处理前下降.miR-7沉默+DADS组较SGC-7901和vector组增殖率下降.miR-7沉默组划痕距离较SGC7901组与vector组明显缩短.DADS 处理后各组细胞的划痕距离较处理前增宽.miR-7沉默组侵袭细胞较SGC7901组与vector组明显增加.DADS处理后各组侵袭细胞较处理前减少,而miR-7沉默+DADS 组减少最为明显.miR-7 沉默组较 SGC7901 组与vector组EGFR、Rac1、Pak1与Vimentin上调和E-cadherin下调.DADS处理后,各组的EGFR、Rac1、Pak1与Vimentin下调和E-cadherin上调,并且miR-7沉默组较各组最为明显.结论 DADS可上调miR-7通过EGFR/Rac1/Pak1信号通路抑制SGC-7901细胞增殖、EMT与迁移侵袭,miR-7沉默起到促进的作用.
Aim To investigate the inhibition of EMT migration and invasion of SGC-7901 cells by DADS down-regulation of miR-7 through EGFR/Rac1/Pak1 pathway and its molecular mechanism.Methods Lentivirus PHBLV-U6-RFP-T2A-Puro-HSA-Mir-7-an-tago and control empty vector pHBLV-U6-RFP-T2A-Puro were transfected with SGC-7901 to establish a stable Mir-7-silenced SGC-7901 cell line.MTT,scratch,migration and invasion assay were used to de-tect cell proliferation,migration and invasion.qRT-PCT,Western blot and immunofluorescence were used to detect the expressions of EGFR,Rac1,Pak1,Vi-mentin and E-cadherin.Results SGC-7901 cells with stable silencing of miR-7 were successfully con-structed.qRT-PCR showed that the expression of miR-7 in SGC7901 transfected with lentivirus si-mir7 was significantly down-regulated.The expression of miR-7 in SGC-7901 cells was significantly up-regulated after DADS treatment.After 24 h,48 h and 72 h,the pro-liferation rate of miR-7 silencing group significantly in-creased compared with SGC7901 group and vector group.The growth rate of DADS after treatment was lower than that before treatment.The proliferation rate of Mir-7-silenced+DADS group was lower than that of SGC-7901 and vector groups.The scratch distance of miR-7 silencing group was significantly shorter than that of SGC7901 group and vector group.The scratch-ing distance of the cells after DADS treatment was wider than that before DADS treatment.The invasion cells of miR-7 silencing group significantly increased compared with SGC7901 group and vector group.The invasion of DADS cells in all groups significantly de-creased after DADS treatment,and the miR-7 silenc-ing+DADS group significantly decreased.EGFR,Rac1,Pak1 and Vimentin were up-regulated and E-cadherin was down-regulated in miR-7 silencing group compared with SGC7901 group and vector group.After DADS treatment,EGFR,Rac1,Pak1 and Vi-mentin were down-regulated and E-cadherin was up-regulated in all groups,with the most significant changes observed in the miR-7 silence group.Conclu-sion DADS can up-regulate miR-7 to inhibit the pro-liferation,EMT,migration and invasion of SGC-7901 cells through EGFR/Rac1/Pak1 pathway,and miR-7 silences play a role in promoting it.
朱欢;邓玲;陈洁海;刘芳;伍尤华;苏琦
湘南学院附属医院肿瘤科,湖南 郴州 423000||湖南省肿瘤细胞与分子病理学重点实验室,肿瘤研究所,湖南 衡阳 421001||南华大学附属南华医院,湖南 衡阳 421000湖南省肿瘤细胞与分子病理学重点实验室,肿瘤研究所,湖南 衡阳 421001||南华大学附属南华医院,湖南 衡阳 421000||湖南省胃癌研究中心,南华大学附属第一医院肿瘤科,湖南 衡阳 421001湖南省肿瘤细胞与分子病理学重点实验室,肿瘤研究所,湖南 衡阳 421001||湖南省胃癌研究中心,南华大学附属第一医院肿瘤科,湖南 衡阳 421001||娄底市中心医院肿瘤科,湖南 娄底 417000湖南省肿瘤细胞与分子病理学重点实验室,肿瘤研究所,湖南 衡阳 421001湖南省胃癌研究中心,南华大学附属第一医院肿瘤科,湖南 衡阳 421001湖南省肿瘤细胞与分子病理学重点实验室,肿瘤研究所,湖南 衡阳 421001||湖南省胃癌研究中心,南华大学附属第一医院肿瘤科,湖南 衡阳 421001
医药卫生
二烯丙基二硫胃癌SGC-7901细胞miR-7EGFR/Rac1/Pak1信号通路EMT迁移侵袭
diallyl disulfidegastric cancer SGC-7901 cellsmiR-7EGFR/Rac1/Pak1 pathEMTmi-gration and invasion
《中国药理学通报》 2026 (4)
644-651,8
国家自然科学基金资助项目(No 81973532)湖南省卫计委科研重点课题(No A2015-2)湖南省高校创新平台开放基金(No 17K081)
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