摘要
目的探讨miRNA-30a-5p(miR-30a-5p)与异黏蛋白(MTDH)体外对人乳腺癌细胞增殖、侵袭、迁移能力的影响。方法利用癌症基因组图谱(TCGA)数据库数据,分析数据库中112例乳腺癌患者癌及癌旁组织MTDH和103例乳腺癌患者癌及癌旁组织miR-30a-5p的表达情况;采用Pearson相关性分析法分析数据库中1222例乳腺癌患者MTDH与miR-30a-5p的相关性;数据更新时间为2022年8月。将乳腺癌MDA-MB-231细胞分为阴性对照组(转染阴性对照序列)、miR-30a-5p过表达组(转染miR-30a-5p模拟物)、siMTDH组[转染针对MTDH基因的小干扰RNA(siMTDH)]、siMTDH+miR-30a-5p过表达组(同时转染siMTDH、miR-30a-5p模拟物);采用四甲基偶氮唑盐(MTT)法检测细胞增殖能力,采用细胞划痕实验检测细胞迁移能力,Transwell法检测细胞侵袭能力,采用实时荧光定量聚合酶链反应(qRT-PCR)检测细胞miR-30a-5p、MTDH、基质金属蛋白酶(MMP)-2、MMP-9、波形蛋白(vimentin)和β-catenin mRNA的相对表达量,采用蛋白质印迹法检测MTDH、N-钙黏蛋白(N-cad)、β-catenin、Snail和MMP-9蛋白的表达。结果TCGA数据库中,乳腺癌组织中MTDH表达水平较癌旁组织高,miR-30a-5p表达水平较癌旁组织低,差异均有统计学意义(均P<0.001);1222例乳腺癌患者MTDH与miR-30a-5p表达呈负相关(r=-0.134,P<0.001)。与阴性对照组比较,siMTDH组和miR-30a-5p过表达组24、48、72h细胞增殖能力均降低(均P<0.001)。miR-30a-5p过表达组、siMTDH组细胞划痕愈合率均低于阴性对照组[(61.6±1.6)%、(54.7±5.9)%比(80.3±3.0)%](均P<0.05)。迁移细胞数均低于阴性对照组[(881±50)个、(725±63)个比(1172±66)个](均P<0.05)。与阴性对照组相比,miR-30a-5p过表达组、siMTDH组MDA-MB-231细胞MMP-2、MMP-9、vimentin和β-catenin mRNA的相对表达量均较阴性对照组下调(均P<0.05)。miR-30a-5p过表达组、siMTDH组MDA-MB-231细胞N-cad、β-catenin、Snail和MMP-9蛋白的相对表达量均较阴性对照组下调(均P<0.05)。
Objective To investigate the effects of miRNA-30a-5p(miR-30a-5p)and metadherin(MTDH)on the proliferation,invasion and migration abilities of human breast cancer cells in vitro.Methods The expression of MTDH in cancer and paracancerous tissues of 112 breast cancer patients in the database and miR-30a-5p in cancer and paracancerous tissues of 103 breast cancer patients in the database were analyzed using data from The Cancer Genome Atlas(TCGA)database.Pearson correlation analysis was used to analyze the correlation between miR-30a-5p and MTDH in 1222 breast cancer patients in the database;the data were updated to August 2022.Breast cancer MDA-MB-231 cells were divided into negative control group(transfected with negative control sequence),miR-30a-5p overexpression group(transfected with miR-30a-5p mimics),siMTDH group[transfected with small interfering RNA against MTDH(siMTDH)],siMTDH+miR-30a-5p overexpression group(transfected with both siMTDH and miR-30a-5p mimics);cell proliferation ability was detected by methyl thiazol tetrazolium(MTT)assay,cell migration ability was detected by cell scratch assay,cell invasion ability was detected by Transwell assay.The relative expressions of miR-30a-5p,MTDH,matrix metalloproteinase(MMP)-2,MMP-9,vimentin andβ-catenin mRNA in cells were detected by quantitative real-time fluorescence polymerase chain reaction(qRT-PCR),and the expressions of MTDH,N-cadherin(N-cad),β-catenin,Snail and MMP-9 proteins were detected by Western blotting.Results In the TCGA database,MTDH expression level was higher and miR-30a-5p expression level was lower in breast cancer tissues compared with paracancerous tissues,and the differences were statistically significant(both P<0.001).There was a negative correlation between MTDH and miR-30a-5p expressions in 1222 patients with breast cancer(r=-0.134,P<0.001).Compared with the negative control group,the cell proliferation ability was reduced in both siMTDH group and miR-30a-5p overexpression group at 24,48 and 72 h(all P<0.001).The cell scratch healing
作者
张晓红
董凡超
周晓
王娟
王志蕙
姚少波
杜阳阳
杨智强
Zhang Xiaohong;Dong Fanchao;Zhou Xiao;Wang Juan;Wang Zhihui;Yao Shaobo;Du Yangyang;Yang Zhiqiang(Department of Pathology,Linyi Tumor Hospital,Linyi 276000,China;Department of Breast Surgery,Linyi Tumor Hospital,Linyi 276000,China;Department of Basic Medicine,Shandong Medical College,Linyi 276000,China;Department of Basic Medicine,Guilin Medical University,Guilin 541004,China)
出处
《肿瘤研究与临床》
CAS
2023年第3期193-199,共7页
Cancer Research and Clinic
基金
临沂市科技发展计划(202120071)。
