Transforming growth factor-β regulates activity of miR-200b-200a-429 promoter

Objective To investigate the regulatory effect of transforming growth factor-β(TGF-β)on miR-200b-200a-429expression and the related mechanism.Methods RT-PCR was used to examine the expression of miR-200a,miR-200band miR-429in HK-2cells treated with TGF-β.The miR-200b-200a-429promoter was amplified by PCR and was cloned into pGL3basic vector,which was then transfected into HK-2cells and the luciferase activity of miR-200b-200a-429gene promoter was detected by luminometer.Results Results of RT-PCR showed that TGF-βdown-regulated miR-200a,miR-200band miR-429in HK-2cells24hafter TGF-βtreatment.Vector of pGL3- miR-200b-200a-429-promoter was constructed successfully and was confirmed by sequencing.Dual-Luciferase reporter gene system was used to verify the promoter activity of the constructed reporter gene vector.TGF-βwas found to significantly inhibit the activities of miR-200b-200a-429promoter in HK-2cells(P0.05).Conclusion TGF-βmay regulate the activity of miR-200b-200a-429promoter.