title: Aberrant expression of microRNAs involved in epithelial-mesenchymal transition of HT-29 cell line.
abstract: Epithelial-mesenchymal transition (EMT) is an essential step for cancer metastasis. MicroRNAs (miRNAs) are small non-coding RNAs that regulate target-mRNAs post-transcriptionally. The expression and function of miRNAs in EMT of HT-29 colonic cells remain elusive. This study looks at expression of miRNAs in EMT and explores the effects of miRNAs on EMT in HT-29 cell line. HT-29 was treated with TGF β to establish an EMT model, in which a collection of miRNAs was dynamically regulated by real-time PCR (qPCR) analysis. Among them, miR-21 and miR-27 were significantly upregulated, while miR-22, miR-26, miR-30, miR-181, miR-200b, miR-200c and miR-214 were markedly downregulated. MiRNA-inhibitors were used to knockdown miRNAs in HT-29 and EMT markers were determined by qPCR to monitor the effects of miRNAs on EMT process. Results showed that miR-22 could not alter the expression of EMT markers, while knockdown of miR-200b could significantly increase that of epithelial markers, N-cadherin, Vimentin, α-Sma and Twist1 and decrease that of mesenchymal marker, E-cadherin. Bioinformatic analysis and Western blot showed that ZEB1 was directly suppressed by miR-200b. In conclusion, miRNAs are dynamically regulated in TGF β-induced EMT of HT-29 and miR-200b was essential for EMT by suppressing the expression of ZEB1 in HT-29.
authors.affiliation: Department of Cardio-Thoracic Surgery, Number 455 Hospital of The Chinese People's Liberation Army, Shanghai 200052, China. caizg12345@yahoo.com.cn
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Cai
authors.forename: Shao-Ming
authors.initials: SM
authors.lastname: Zhang
authors.forename: Hang
authors.initials: H
authors.lastname: Zhang
authors.forename: Yi-Yong
authors.initials: YY
authors.lastname: Zhou
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Wu
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Xu
mesh_terms: D020413:3' Untranslated Regions; D000199:Actins / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D019295:Computational Biology; D015536:Down-Regulation / Q000187:drug effects; D058750:Epithelial-Mesenchymal Transition; D019073:HT29 Cells; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D051879:Twist-Related Protein 1 / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D020413:3' Untranslated Regions; C541116:ACTA2 protein, human; D000199:Actins; D015820:Cadherins; D018398:Homeodomain Proteins; D035683:MicroRNAs; D009687:Nuclear Proteins; C103173:TWIST1 protein, human; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D051879:Twist-Related Protein 1; D014746:Vimentin; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1