miR-153-5p靶向基质金属蛋白酶-16对胃癌细胞增殖、迁移及侵袭的影响  

Effect of miR-153-5p targeting matrix metalloproteinase-16 on proliferation,migration and invasion of gastric cancer cells

作　　者：胡小敏 王燕 程旭敏 罗寰[1] HU Xiao-min;WANG Yan;CHENG Xu-min;LUO Huan(Department of Gastrointestinal Oncology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]河北北方学院附属第一医院胃肠肿瘤外科,河北张家口075000

出　　处：《中国生物制品学杂志》2022年第12期1449-1457,共9页Chinese Journal of Biologicals

基　　金：河北省医学科学研究课题(20191648)。

摘　　要：目的探讨miR-153-5p对胃癌细胞SGC-7901增殖、迁移和侵袭的影响。方法RT-qPCR法检测胃癌和癌旁组织(河北北方学院附属第一医院2016年1月—2018年12月经病理确诊的胃癌患者手术切除样本)以及正常胃细胞GES-1和胃癌细胞系(GES-1、MHCC97H、MKN-1、SGC-7901、MKN45、MGC803)中miR-153-5p的表达;将SGC-7901细胞分别或联合转染各质粒后分为miR-NC、miR-153-5p mimic、miR-153-5p inhibitor、pc-NC、pc-MMP-16、miR-153-5p mimic+pc-NC和miR-153-5p mimic+pc-MMP-16组,以未处理的SGC-7901细胞为对照组。克隆形成试验检测细胞增殖能力,Transwell试验检测细胞侵袭能力,划痕试验检测细胞迁移能力,Western blot法检测基质金属蛋白酶-16(matrix metalloproteinase-16,MMP-16)表达水平,Targetscan网站和双荧光素酶报告试验分别预测和验证miR-153-5p与MMP-16的靶向关系。结果在胃癌中,miR-153-5p的表达与性别、年龄和吸烟史无关(χ^(2)分别为0.001、1.1002、0.096,P均>0.05),而与肿瘤大小、肿瘤-淋巴结-远处转移(tumor-node-metastasis,TNM)分期、淋巴结转移及分化成度有关(χ^(2)分别为4.444、5.094、8.487、9.427,P均<0.05);miR-153-5p在胃癌组织和细胞中低表达。与对照组相比,miR-153-5p mimic组细胞克隆形成率、划痕闭合率及侵袭细胞数均显著降低(t分别为7.744、4.199、6.423,P均<0.05);miR-153-5p与MMP-16具有靶向关系,在3′UTR区存在结合位点;与miR-NC组相比,miR-153-5p mimic组MMP-16蛋白表达水平显著下调(t=11.532,P<0.01),与pc-NC组相比,pc-MMP-16组MMP-16蛋白表达水平显著上调(t=10.694,P<0.01);与miR-153-5p mimic+pc-NC组相比,miR-153-5p mimic+pc-MMP-16组细胞MMP-16蛋白表达水平、细胞克隆形成率、细胞侵袭数、划痕闭合率均显著升高(t分别为13.802、7.762、9.752、5.091,P均<0.01)。结论miR-153-5p通过靶向MMP-16抑制胃癌细胞增殖、侵袭和迁移。Objective To investigate the effect of miR-153-5p on the proliferation,migration and invasion of gastric cancer SGC-7901 cells.Methods The expressions of miR-153-5p in gastric cancer and adjacent tissues(collected from surgically resected samples of patients with gastric cancer pathologically diagnosed from January 2016 to December 2018 in The First Affiliated Hospital of Hebei North University),normal gastric GES-1 cells and gastric cancer cell lines(GES-1,MHCC97H,MKN-1,SGC-7901,MKN45 and MGC803)were determined by RT-qPCR.SGC-7901 cells were divided into seven groups and transfected with miR-NC,miR-153-5p mimic,miR-153-5p inhibitor,pc-NC,pc-MMP-16,miR-153-5p mimic+pc-NC and miR-153-5p mimic+pc-MMP-16 respectively,using those untreated as control,and determined for proliferation ability by clone formation test,for invasion ability by Transwell assay,and for migration ability by scratch test.The expression levels of matrix metalloproteinase-16(MMP-16)was determined by Western blot,while the targeting relationship between miR-153-5p and MMP-16 were predicted and validated by Targetscan website and dual luciferase reporter assay respectively.Results The expression of miR-153-5p in gastric cancer was not related to the gender,age and smoking history(χ^(2)=0.001,1.1002 and 0.096 respectively,each P>0.05),while was related to the tumor size,tumornode-metastasis(TNM)stage,lymph node metastasis and degree of differentiation(χ^(2)=4.444,5.094,8.487 and 9.427respectively,each P<0.05);miR-153-5p was low expressed in gastric cancer tissues and cells.Compared with those in control group,the cell clone formation rate,scratch closure rate and account of invasive cells in miR-153-5p mimic group decreased significantly(t=7.744,4.199 and 6.423 respectively,each P<0.05);miR-153-5p showed a targeting relationship with MMP-16 with a binding site in the 3′UTR region.The expression level of MMP-16 protein in miR-153-5p mimic group decreased significantly(t=11.532,P<0.01)as compared with that in miR-NC group,while that in pcMMP-16 g

关 键 词：miR-153-5p 基质金属蛋白酶-16 胃癌细胞 增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]