摘要
目的:探讨miR-144-5p在甲状腺乳头状癌(papillary thyroid carcinoma, PTC)细胞中的表达趋势及对PTC细胞系增殖、侵袭及迁移能力的影响及相关分子机制研究。方法:实时荧光定量聚合酶链反应(quantitative real-time PCR,qRT-PCR)检测人PTC细胞系TPC-1、 BCPAP及人正常甲状腺滤泡细胞系Nthy-ori 3-1中miR-144-5p的表达情况。将miR-144-5p-mimics、miR-144-5p-inhibitor及inhibitor-NC和mimics-NC分别以Lipofectamin^(TM)2000转染至TPC-1细胞。分为4组:阴性对照组(mimics-NC组)和过表达组(mimics-144-5p组)。阴性对照组(inhibitor-NC组)和抑制组(inhibitor-144-5p组)。以CCK-8生长曲线、小室侵袭实验和划痕实验检测4组细胞增殖、侵袭和迁移能力。ENCORI数据库验证miR-144-5p在TCGA中的表达趋势。在线数据库预测miR-144-5p下游靶基因。通过STING得到靶基因蛋白互作网络图。通过Cytascap插件CytoHubba得Degree值为Top10的基因。GEPIA数据库检测ACTG1和STAT1在TCGA数据库中的表达趋势。HPA数据库验证ACTG1和STAT1蛋白表达水平。qRT-PCR和TaretScan在线数据库验证miR-144-5p与ACTG1和STAT1之间的调控关系。结果:PTC细胞系中miR-144-5p表达趋势明显低于Nthy-ori 3-1人正常甲状腺滤泡上皮细胞。上调miR-144-5p后TPC-1细胞的增殖活性、侵袭和迁移能力明显下降,而抑制miR-144-5p则起到相反的作用。在线数据库预测miR-144-5p下游靶基因为584个。通过STING得到靶基因蛋白互作网络图。通过Cytascap插件CytoHubba得Degree值为Top10的基因。在线数据库ENCORI验证ACTG1和STAT1可能是miR-144-5p的下游靶基因。qRT-PCR验证miR-144-5p与ACTG1和STAT1之间存在负调控关系。结论:miR-144-5p在PTC细胞中低表达,过表达或敲低miR-144-5p可能是通过调控ACTG1和STAT1影响PTC细胞的增殖、侵袭及迁移。
Objective:To investigate the expression trend of miR-144-5p in papillary thyroid carcinoma(PTC)cells and its effect on the proliferation,invasion and migration of PTC cell lines and related molecular mechanisms.Methods:Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-144-5p in human PTC cell lines TPC-1 and BCPAP,and human normal thyroid follicular cell line Nthy-ori 3-1.miR-144-5p-mimics,miR-144-5p-inhibitor,inhibitor-NC and mimics-NC were transfected into TPC-1 cells by Lipofectamin^(TM) 2000,respectively.The cells were divided into 4 groups:Negative control group(mimics-NC group),overexpression group(mimics-144-5p group),negative control group(inhibitor NC group)and inhibitor-144-5p group(inhibitor-144-5p group).The cell proliferation,invasion and migration of the four groups were detected by CCK-8 growth curve,chamber invasion test and scratch test.ENCORI database was used to verify the expression trend of miR-144-5p in TCGA.The downstream target genes of miR-144-5p were predicted by online database.The target gene-protein interaction network was obtained by STING.The genes with Degree value Top10 were obtained by Cytascap plugin CytoHubba.GEPIA database was used to detect the expression trends of ACTG1 and STAT1 in TCGA database.ACTG1 and STAT1 protein expression levels were verified by HPA database.qRT-PCR and TaretScan online database were used to verify the regulatory relationship between miR-144-5p and ACTG1 and STAT1. Results: The expression trend of miR-144-5p in papillary thyroid carcinoma(PTC) cell lines was significantly lower than that in human normal thyroid follicular epithelium cell line Nthy-ori 3-1.The proliferation activity,invasion and migration ability of TPC-1 cells were significantly decreased after the upregulation of miR-144-5p,while the inhibition of miR-144-5p had the opposite effect.The online database predicted 584 downstream target genes of miR-144-5p.The target gene-protein interaction network was obtained by STING.The genes with De
作者
高圣钰
丁成龙
魏微微
GAO Shengyu;DING Chenglong;WEI Weiwei(Department of General Surgery,the First Affiliated Hospital of Jiamusi University,Heilongjiang Jiamusi 154002,China;Department of Pathology,the First Affiliated Hospital of Jiamusi University,Heilongjiang Jiamusi 154002,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第9期1595-1601,共7页
Journal of Modern Oncology
基金
黑龙江省卫生健康委课题(编号:2020-328)。
