Hsa-miR-5581-3p and hsa-miR-542-3p target the F8 gene in hemophilia A without F8 mutations F8 gene is regulated by miRNAs

Main Article Content

Feiying Meng

Keywords

Hemophilia A, Hsa-miR-5581-3p, Hsa-miR-542-3p, F8 gene

Abstract

Objective: This study aims at uncovering the effects of microRNAs (miRNAs) on F8 gene and FVIII protein in hemophilia A (HA).


Methods: F8-targeting miRNAs were predicted by TargetScan, miRDB and starBase. MiRNAs predicted by at least two of the three databases were selected for further study, and their expressions in the blood of HA patients without F8 mutations and healthy controls were detected. Dual-luciferase reporter assay was performed to verify the binding between hsa-miR-5581-3p/hsa-miR-542-3p and F8. The regulation of F8 by hsa-miR-5581-3p/hsa-miR-542-3p was investigated in human umbilical vein endothelial cells (HUVECs) and lymphoblastoid cell line (LCL) that displayed endogenous expression of FVIII. qRT-PCR was used to detect the expressions of miRNAs and F8 gene, and Western blotting was conducted to measure the expression of FVIII protein.


Results: A total of 43 F8-targeting miRNAs were predicted by at least two of the three databases. Among these miRNAs, hsa-miR-5581-3p and hsa-miR-542-3p were highly expressed in the blood of HA patients and have not been reported in previous studies of HA. Both hsa-miR-5581-3p and hsa-miR-542-3p could bind the 3’UTR of F8 mRNA. Upregulation of hsa-miR-5581-3p or hsa-miR-542-3p suppressed the expressions of F8 mRNA and FVIII protein in HUVECs and LCL cells.


Conclusion: Hsa-miR-5581-3p and hsa-miR-542-3p target the F8 gene and suppress the expression of FVIII protein, which may contribute to the development of HA without F8 mutations.

Downloads

Download data is not yet available.
Abstract 159 | PDF Downloads 49 HTML Downloads 11