MiR-125b-5p and miR-181b-5p inhibit keratinocyte proliferation in skin by targeting Akt3

Abstract

MicroRNAs (miRNAs) have been widely accepted to play important roles in the regulation of keratinocyte functions. Here, we aimed to further explore the role and underlying mechanism of miR-125b-5p and miR-181b-5p in psoriasis. The expression levels of miR-125b-5p, miR-181b-5p and Akt3 mRNA were detected by qRT-PCR assay. Cell proliferation ability was determined by MTT assay and BrdU incorporation assay. Dual-luciferase reporter assay and RNA Immunoprecipitation assay were used to confirm the targeted interaction between miR-125b-5p or miR-181b-5p and Akt3 in human epidermal keratinocytes (HEKs). The levels of ki-67, Akt3 protein, Akt, p-Akt, mTOR and p-mTOR were measured by Western blot. Our study indicated that miR-125b-5p and miR-181b-5p were downregulated (about 61.3% with miR-125b-5p and 60.4% with miR-181b-5p) and Akt3 was upregulated (about 2.68-fold) in psoriasis. Upregulation of miR-125b-5p or miR-181b-5p resulted in about a 33% or 40% reduction of HEKs proliferation in vitro, while Akt3 overexpression triggered a 1.3-fold enhancement on HEKs proliferation. Akt3 was a direct target of miR-125b-5p or miR-181b-5p. Moreover, HEKs proliferation ability in cotransfection of miR-125b-5p mimics (or miR-181-5p mimics) and vector-Akt3 group was about 2-fold (or 1.98-fold) that in the miR-125b-5p mimics (or miR-181-5p mimics) alone group. Akt/mTOR signaling was involved in miR-125b-5p mimics- or miR-181b-5p mimics-mediated inhibition effect on HEKs proliferation. Our study suggested that the upregulation of miR-125b-5p or miR-181b-5p inhibited HEKs proliferation at least partly by targeting Akt3, providing novel mechanisms of miRNAs involved in psoriasis.

Keywords: Human epidermal keratinocytes (HEKs); Proliferation; Psoriasis; miR-125b-5p; miR-181b-5p.