miR-875-5p通过靶向USF2抑制胃癌细胞的增殖、迁移和侵袭  

miR-875-5p inhibits the proliferation, migration and invasion of gastric cancer cells by targeting USF2

作　　者：高慎硕 张智凯 尹国庆 张红霞[1] 王绪斌 马岩[1] 刘洪俊[1] 李乐平[1] 郭晓波[1] GAO Shenshuo;ZHANG Zhikai;YIN Guoqing;ZHANG Hongxia;WANG Xubin;MA Yan;LIU Hongjun;LI Leping;GUO Xiaobo(Department of Gastrointestinal Surgery,Shandong Provincial Hospital Affiliated to Shandong University,Jinan 250021,Shandong,China;Department of Gastrointestinal Surgery,Qingzhou Hospital Affiliated to Shandong First Medical University,Qingzhou 262500,Shandong,China;Clinical Mecical College,Shandong First Medical University,Jinan 250117,Shandong,China)

机构地区：[1]山东大学附属省立医院胃肠外科,山东济南250021 [2]山东第一医科大学附属青州医院胃肠外科,山东青州262500 [3]山东第一医科大学临床医学院,山东济南250117

出　　处：《中国肿瘤生物治疗杂志》2022年第1期23-29,共7页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81672379)。

摘　　要：目的:探讨miR-875-5p对胃癌细胞增殖、迁移和侵袭的影响及其机制。方法:采用qPCR法检测胃癌细胞BGC-823、HGC-27、MGC-803、SGC-7901、AGS、MKN-45和胃黏膜上皮细胞GES-1中miR-875-5p的表达水平。利用脂质体转染技术,分别将miR-875-5p模拟物/抑制剂(mimic/inhibitor)及其阴性对照质粒(miR-NC/Anti-miR-NC)转染至AGS细胞/MKN-45细胞,构建过表达/抑制miR-875-5p的细胞模型,空白对照组(Control组)不转染。通过CCK-8、克隆形成、Transwell等实验分别检测miR-875-5p表达变化对细胞增殖、克隆形成、迁移和侵袭的影响。采用双荧光素酶报告基因实验验证miR-875-5p与上游刺激因子2(USF2)的靶向关系,WB实验验证miR-875-5p对USF2的调控作用并检测USF2蛋白的表达。构建MKN-45细胞裸鼠移植瘤模型,验证miR-875-5p过表达对MKN-45细胞成瘤能力的影响。结果:miR-875-5p在6种胃癌细胞中表达水平显著低于胃黏膜上皮细胞GES-1(均P<0.01)。与Control组和miR-NC组相比,miR-875-5p mimic组AGS细胞的增殖、克隆形成率、迁移和侵袭细胞数,以及USF2蛋白的表达均显著降低(P<0.05或P<0.01);miR-875-5p inhibitor组MKN-45细胞的增殖、克隆形成率、迁移和侵袭细胞数,以及USF2蛋白的表达均显著提高(P<0.05或P<0.01)。双荧光素酶报告基因实验证明,miR-875-5p能够直接靶向USF2基因。体内成瘤实验结果表明,过表达miR-875-5p显著抑制MKN-45细胞移植瘤的生长(均P<0.01)。结论:miR-875-5p通过靶向USF2抑制胃癌细胞的增殖、迁移和侵袭。Objective: To investigate the effects of miR-875-5 p on proliferation, migration and invasion of gastric cancer(GC) cells and its mechanism. Methods: The expression level of miR-875-5 p was detected by qPCR in GC cell lines(BGC-823, HGC-27,MGC-803, SGC-7901, AGS, MKN-45) and gastric epithelial GES-1 cells. MiR-875-5 p mimics/inhibitors or their negative control plasmids(miR-NC/anti-miR-NC) were transfected into AGS or MKN-45 cells by liposome transfection technique to construct miR-875-5 p overexpression or knockdown cell model. Cells in blank control group(Control group) were not transfected. The effects of miR-875-5 p on cell proliferation, clone formation, migration and invasion were detected by CCK-8 assay, colony formation assay and Transwell assay, respectively. The targeting relationship between miR-875-5 p and upstream stimulatory factor 2(USF2) was verified by dual-luciferase reporter gene assay. The regulation of miR-875-5 p on USF2 as well as the expression of USF2 protein was confirmed by WB assay. After the construction of MKN-45 cell transplanted tumor model in nude mice, the effect of miR-875-5 p overexpression on tumorigenesis of MKN-45 cells was detected. Results: The expression level of miR-875-5 p in 6 GC cells was significantly lower than that in gastric epithelial GES-1 cells(all P<0.01). Compared with Control group and miR-NC group, the proliferation, clone formation rate, number of migrated and invaded cells, and USF2 protein expression level in AGS cells were significantly decreased in miR-875-5 p mimic group(P<0.05 or P<0.01), while those were significantly increased in MKN-45 cells of miR-875-5 p inhibitor group(P<0.05 or P<0.01). Dual-luciferase reporter gene assay demonstrated that miR-875-5 p could directly target USF2 gene. In vivo tumorigenesis experiment results showed that overexpression of miR-875-5 p significantly inhibited the growth of MKN-45 cell transplanted tumors(all P<0.01). Conclusion: miR-875-5 p inhibits proliferation, migration and invasion of GC cells by targeting US

关 键 词：胃癌 AGS细胞 MKN-45细胞 mi R-875-5p 上游刺激因子2 增殖 迁移 侵袭 

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