miR-153-3p通过靶向FZD3调控胃癌SGC7901细胞的增殖、侵袭与迁移  

miR-153-3p regulates proliferation,invasion and migration of gastric cancer SGC7901 cells via targeting FZD3

作　　者：张曹[1] 何亚琴[2] 钱海权 叶晶晶 ZHANG Cao;HE Yaqin;QIAN Haiquan;YE Jingjing(Department of Gastrointestinal Surgery,the General Hospital of Ningxia Medical University,Yinchuan 750001,Ningxia,China;Department of Surgery,the General Hospital of Ningxia Medical University,Yinchuan 750001,Ningxia,China;Function Inspection Department of Heart Center,the General Hospital of Ningxia Medical University,Yinchuan 750001,Ningxia,China)

机构地区：[1]宁夏医科大学总医院胃肠外科,宁夏银川750001 [2]宁夏医科大学总医院外科学研究室,宁夏银川750001 [3]宁夏医科大学总医院心脏中心功能检查部,宁夏银川750001

出　　处：《中国肿瘤生物治疗杂志》2021年第9期885-892,共8页Chinese Journal of Cancer Biotherapy

基　　金：宁夏自然科学基金资助项目(No.2019AAC03200,No.2019AAC03225)。

摘　　要：目的:探讨miR-153-3p对胃癌SGC7901细胞增殖、侵袭和迁移的作用及其机制。方法:收集2018年5月至2020年6月宁夏医科大学总医院收治的60例胃癌患者的癌和配对癌旁组织标本,以及人胃癌细胞系NCI-N87、AGS、SNU-5、SGC7901和胃上皮细胞GES-1,qPCR法检测miR-153-3p在胃癌组织与细胞中的表达水平。将miR-153-3p mimic及mimic对照序列转染至SGC7901细胞,用CCK-8、克隆形成、流式细胞术、TUNEL、Transwell和划痕愈合实验分别检测上调miR-153-3p对SGC7901细胞增殖、凋亡、侵袭和迁移的影响。构建裸鼠SGC7901细胞移植瘤模型,观察miR-153-3p对肿瘤生长的影响。通过生物信息学数据库和双荧光素酶报告基因实验预测并验证miR-153-3p与FZD3靶向关系,WB法检测miR-153-3p对FZD3蛋白及Wnt/β-catenin通路相关蛋白表达的影响。结果:miR-153-3p在胃癌组织和细胞中表达水平分别显著低于癌旁组织和GES-1细胞(均P<0.01),以SGC7901细胞中表达水平最低。上调miR-153-3p显著抑制SGC7901细胞的增殖、侵袭和迁移能力,并提高细胞凋亡率(均P<0.01),同时上调细胞中E-cadherin表达而下调N-cadherin、MMP2和MMP9表达(均P<0.01)。在体内实验表明,静脉注射miR-153-3p mimic显著降低移植瘤体积和瘤组织中Ki-67表达而上调P57表达(均P<0.01)。机制分析表明,miR-153-3p靶向结合FZD3基因的3′UTR区域,上调miR-153-3p会抑制FZD3表达并上调β-catenin、TCF-4和cyclin D1水平(均P<0.01)。结论:miR-153-3p靶向FZD3并通过Wnt/β-catenin信号通路调控胃癌SGC7901细胞的增殖、侵袭和迁移。Objective:To investigate the effect of miR-153-3p on the proliferation,invasion and migration of gastric cancer SGC7901 cells and its molecular mechanism.Methods:A total of 60 pairs of cancer tissues and corresponding para-cancerous tissues of gastric cancer patients who were treated in the General Hospital of Ningxia Medical University during May 2018 and June 2020 were collected for this study;in addition,human gastric cancer cell lines(NCI-N87,AGS,SNU-5,SGC7901)and normal gastric epithelial GES-1 cells were also collected.The expression level of miR-153-3p in gastric cancer tissues and cells was detected by qRT-PCR.miR-153-3p mimics and mimic control were transfected into SGC7901 cells,respectively.The effects of miR-153-3p overexpression on proliferation,apoptosis,invasion and migration of gastric cancer SGC7901 cells were respectively determined using CCK-8,Clone formation assay,Flow cytometry,TUNEL,Transwell and Wound-healing assay.SGC7901 cell transplanted xenograft tumor model in nude mice was established to explore the effect of miR-153-3p on tumor growth.The targeting relationship between miR-153-3p and FZD3 was predicted and verified by bioinformatics database and Dual-luciferase reporter gene assay,respectively.The effects of miR-153-3p on expression of FZD3 protein and Wnt/β-catenin pathway related proteins were determined by Western blotting.Results:The expression level of miR-153-3p in gastric cancer tissues and cells was significantly lower than that in para-cancerous tissues and gastric epithelial cells,with the lowest expression in SGC7901 cells(all P<0.01).Up-regulation of miR-153-3p significantly inhibited the proliferation,invasion and migration abilities of SGC7901 cells,and increased the rate of apoptotic cells(all P<0.01).In addition,up-regulation of miR-153-3p significantly promoted the expression of E-cadherin but significantly suppressed the expression of N-cadherin,matrix metalloproteinase-2(MMP2)and matrix metalloproteinase-9(MMP9)(all P<0.01).In vivo experiments showed that intraven

关 键 词：miR-153-3p FZD3基因 胃癌 SGC7901细胞 增殖 侵袭 迁移 WNT/Β-CATENIN信号通路 

分 类 号：R735.2[医药卫生—肿瘤] R730.2[医药卫生—临床医学]