Transcription factor CCAAT/enhancer-binding protein-ß upregulates microRNA, let-7f-1 in human endocervical cells.
Am J Reprod Immunol
; 78(6)2017 Dec.
Article
em En
| MEDLINE
| ID: mdl-28921745
ABSTRACT
PROBLEM:
In endocervical epithelial cells (End1/E6E7), miRNA let-7f plays an important role in the control of innate immunity. The underlying molecular mechanism for let-7f regulation in these cells remains largely unclear. METHODS OF STUDY let-7f was knocked down in End1/E6E7 cells using siRNA, and differential gene expression was analyzed by microarray. Differentially expressed genes were validated by qPCR and Western blot. Expression of let-7f was studied by qPCR after inhibition of C/EBPß with betulinic acid (BA) and pCMVß plasmid and after overexpression of C/EBPß with pCMVß+ plasmid. ChIP assay was performed to confirm binding of C/EBPß to let-7f promoter. Levels of Lin28A/B were checked by qPCR after similar treatment.RESULTS:
let-7f knockdown (KD) affects the expression of many transcription factors (eg, C/EBPß) which are important regulators of immune responses. We observed let-7f-1 promoter to contain 6 C/EBPß binding sites. KD of C/EBPß led to decreased let-7f expression while overexpression of C/EBPß increased its expression. Treatment of End1/E6E7 cells with TLR-3 ligand, poly(IC) increased binding of C/EBPß at binding sites 3, 5, and 6. Expression of Lin28A/B also changed upon inhibition and overexpression of C/EBPß. Its expression is opposite to that of let-7f in End1/E6E7 cells.CONCLUSION:
let-7f-1 is a direct target of transcription factor, C/EBPß in End1/E6E7 cells.Palavras-chave
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Proteína beta Intensificadora de Ligação a CCAAT
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MicroRNAs
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Células Epiteliais
Limite:
Female
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Humans
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article