title: MiR-200 Regulates Epithelial-Mesenchymal Transition in Anaplastic Thyroid Cancer via EGF/EGFR Signaling.
abstract: This study was set to study the molecular mechanism underlying how miR-200 regulates EGF/EGFR signaling to involve in epithelial-mesenchymal transition (EMT) in anaplastic thyroid cancer (ATC) cells. Loss-of-function experiments of EGFR silencing by siRNA transfection was performed. Transfection of pre-miR-200s or anti-miR-200s was used to increase or decrease miR-200 transcripts. Real-time PCR, Western blot, immunohistochemistry, and transwell experiments were performed to determine the role of miR-200s in EMT and its role in EGF/EGFR-mediated EMT in vitro and in vivo. EGF/EGFR signaling activation increased the expression of mesenchymal marker vimentin in Nthy-ori 3-1 cells and decreased the expression of endothelial maker E-cadherin. EGF stimulation led to increased RhoA expression in Nthy-ori 3-1 cells. EGFR silencing resulted in decreased RhoA expression in SW1736 and ARO cells. EGF stimulation led to down-regulation of miR-200s and EMT. Restoration of miR-200 expression by pre-miR-200a/c transfection reversed the process, including increased E-cadherin and decreased vimentin. Down-regulation of miR-200 by anti-miR-200 effectively reduced miR-200. Matrigel invasion assay proved that restoration of miR-200 expression counteracted invasiveness. EGFR silencing decreased invasiveness in SW1736 cells, while down-regulation of miR-200s restored invasiveness. Xenograft tumors of SW1736 cells with cotransfection of anti-miR-200s and EGFR siRNA which kept the similar E-cadherin and vimentin expression with the untransfected controls. In ATC cells, miR-200s play a central role in EGF/EGFR-mediated invasiveness in vitro and EMT in vivo.
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xue
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China.
authors.forename: Dongyue
authors.initials: D
authors.lastname: Su
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China.
authors.forename: Rongrong
authors.initials: R
authors.lastname: Yuan
authors.affiliation: Department of Endocrinology, Henan University Huaihe Hospital, Kaifeng, 475000, China. wuyan_p@163.com.
authors.forename: Wuyan
authors.initials: W
authors.lastname: Pang
mesh_terms: D015703:Antigens, CD; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D015536:Down-Regulation; D004815:Epidermal Growth Factor / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition; D066246:ErbB Receptors / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D065646:Thyroid Carcinoma, Anaplastic / Q000378:metabolism*; D013964:Thyroid Neoplasms / Q000378:metabolism*; D014746:Vimentin / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014746:Vimentin; C066737:RHOA protein, human; D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors; D020742:rhoA GTP-Binding Protein
keywords: Anaplastic thyroid cancer; EGF/EGF; Epithelial–mesenchymal transition; Molecular mechanism; miR-200