miR-100-5p调控甲状腺乳头状癌细胞的增殖、迁移、侵袭及EMT进程  

miR-100-5p regulates the proliferation,migration,invasion and EMT process of thyroid papillary carcinoma cells

作　　者：魏微微[1] 高圣钰 孟凡石[1] WEI Weiwei;GAO Shengyu;MENG Fanshi(Department of General Surgery,First Affiliated Hospital of Jiamusi University,Heilongjiang Jiamusi 154002,China)

机构地区：[1]佳木斯大学附属第一医院普外科,黑龙江佳木斯154002

出　　处：《现代肿瘤医学》2023年第22期4123-4129,共7页Journal of Modern Oncology

基　　金：黑龙江省卫生健康委课题(编号:2020-328)。

摘　　要：目的:探讨miR-100-5p对甲状腺乳头状癌细胞增殖、迁移、侵袭及上皮间质转化(EMT)进程的影响。方法:通过GEO数据库下的miRNA微阵列数据集(GSE104006、GSE73182、GSE151180),选取变化倍数[|log2Fold Change(FC)|>1]和adj.P.Val<0.05的miRNA进行交集。实时荧光定量PCR(qRT-PCR)检测miR-100-5p在PTC细胞系(TPC-1和B-CPAP)和人正常甲状腺细胞系(Nthy-ori 3-1)中的表达趋势。合成miR-100-5p相关的模拟物(mimics-100-5p)、抑制剂(inhibitor-100-5p)以及对应的对照组(mimics-NC和inhibitor-NC)。分别转染PTC细胞系,通过Edu实验、平板克隆实验、划痕愈合实验、Transwell实验深入研究miR-100-5p对PTC细胞的影响。通过蛋白印迹实验检测转染mimics-100-5p、inhibitor-100-5p及对应NC组MMP-9和EMT相关标志蛋白的表达趋势。结果:三个miRNA微阵列数据库共同交集于miR-100-5p。与人正常甲状腺细胞系Nthy-ori 3-1相比,miR-100-5p在PTC细胞系中低表达。qRT-PCR验证转染成功。Edu实验、平板克隆实验显示过表达miR-100-5p细胞的增殖速率明显降低,而敲低miR-100-5p后结果与之相反。划痕愈合实验和Transwell实验显示过表达miR-100-5p的TPC-1细胞的迁移和侵袭能力受限,而敲低miR-100-5p的结果与之相反。蛋白印记实验显示过表达miR-100-5p后MMP-9、VIM蛋白表达趋势明显降低,而E-cad蛋白表达上升,敲低miR-100-5p可以逆转过表达miR-100-5p的结果。结论:miR-100-5p负调控PTC细胞的增殖、迁移和侵袭。此外,miR-100-5p还参与介导PTC的上皮间质转化进程。Objective:To investigate the effect of miR-100-5p on the proliferation,migration,invasion and EMT process of papillary thyroid carcinoma cells.Methods:Through the miRNA microarray datasets(GSE104006,GSE73182,GSE151180) under the GEO database,the miRNAs with the fold change [|log2Fold Change(FC)| 1]and adj.P.Val 0.05 were selected for intersection.Real-time fluorescent quantitative PCR(qRT-PCR) was used to detect the expression trend of miR-100-5p in PTC cell lines(TPC-1 and B-CPAP) and human normal thyroid cell line(Nthy-ori 3-1).Synthesize miR-100-5p-related mimics(mimics-100-5p),inhibitors(inhibitor-100-5p) and corresponding control groups(mimics-NC and inhibitor-NC).PTC cell lines were transfected,and the effects of miR-100-5p on PTC cells were studied in depth by Edu experiment,plate cloning experiment,scratch healing experiment and Transwell experiment.The expression trends of MMP-9 and EMT-related marker proteins in the transfected mimics-100-5p,inhibitor-100-5p and corresponding NC groups were detected by Western blot experiments.Results:Three miRNA microarray databases intersected in miR-100-5p.Compared with human normal thyroid cell line Nthy-ori 3-1,miR-100-5p was lower expressed in PTC cell line.qRT-PCR verified that the transfection was successful.Edu experiment and plate cloning experiment showed that the proliferation rate of cells overexpressing miR-100-5p was significantly reduced,but the results were opposite after knocking down miR-100-5p.Scratch healing assay and Transwell assay showed that miR-100-5p overexpressed TPC-1cells had limited migration and invasion abilities,while the results of knocking down miR-100-5p were contrary.Western blot experiments showed that after overexpressing miR-100-5p,the protein expression of MMP-9 and VIM decreased significantly,while the expression of E-cad protein increased.Knocking down miR-100-5p could reverse the result of overexpressing miR-100-5p.Conclusion:miR-100-5p negatively regulates the proliferation,migration and invasion of PTC.cells.In addition,miR

关 键 词：miR-100-5p 甲状腺乳头状癌 EMT 

分 类 号：R736.1[医药卫生—肿瘤]