Specificity Protein 1 ( Sp1) Maintains Basal Epithelial Expression of the miR-200 Family

Background: Epithelial-mesenchymal transition (EMT) is a key process in embryonic development and cancer metastasis. Results: Sp1 activates miR-200 transcription in epithelial cells and prevents EMT. Conclusion: miR-200 family members require Sp1 to drive basal expression and maintain an epithelial state. Significance: Defining the mechanisms controlling the epithelial state has implications for understanding early differentiation and for designing interventions to prevent cancer metastasis. Epithelial-mesenchymal transition (EMT) is required for the specification of tissues during embryonic development and is recapitulated during the metastatic progression of tumors. The miR-200 family plays a critical role in enforcing the epithelial state with their expression lost in cells undergoing EMT. EMT can be mediated by activation of the ZEB1 and ZEB2 (ZEB) transcription factors, which repress miR-200 expression via a self-reinforcing double negative feedback loop to promote the mesenchymal state. However, it remains unclear what factors drive and maintain epithelial-specific expression of miR-200 in the absence of EMT-inducing factors. Here, we show that the transcription factor Specificity Protein 1 (Sp1) binds to the miR-200b approximate to 200a approximate to 429 proximal promoter and activates miR-200 expression in epithelial cells. In mesenchymal cells, Sp1 expression is maintained, but its ability to activate the miR-200 promoter is perturbed by ZEB-mediated repression. Reduction of Sp1 expression caused changes in EMT-associated markers in epithelial cells. Furthermore, we observed co-expression of Sp1 and miR-200 during mouse embryonic development wherein miR-200 expression was only lost in regions with high ZEB expression. Together, these findings indicate that miR-200 family members require Sp1 to drive basal expression and to maintain an epithelial state.