Abstract
Keratinocyte migration is a crucial process during skin wound healing, and circular RNAs are associated with keratinocyte migration. The purpose of our study was to clarify the role of circ_0084443 in wound healing. The levels of circ_0084443, microRNA (miR)-17-3p, and forkhead box protein O4 (FOXO4) were examined by quantitative reverse transcription-PCR. Cell migration was detected via wound scratch assay or transwell assay. The protein expression was measured using western blot. The binding analysis between miR-17-3p and circ_0084443 or FOXO4 was determined by dual-luciferase reporter assay and RNA Immunoprecipitation assay. TGF-β1 decreased the levels of circ_0084443 and FOXO4 while increased the miR-17-3p expression in keratinocytes by a concentration-dependent manner. Circ_0084443 acted as a miR-17-3p sponge and circ_0084443 overexpression alleviated TGF-β1-induced migration of keratinocytes by sponging miR-17-3p. FOXO4 was a target for miR-17-3p. The downregulation of miR-17-3p suppressed cell migration in TGF-β1-induced cells by increasing the FOXO4 level. Circ_0084443 positively regulated the FOXO4 expression by sponging miR-17-3p. Circ_0084443 suppressed the TGFβ signaling pathway by affecting the miR-17-3p/FOXO4 axis. These results exhibited that circ_0084443 suppressed the TGF-β1-induced keratinocyte migration by regulating the miR-17-3p/FOXO4 axis, suggesting the application potential of circ_0084443 in wound-healing-related diseases.
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The analyzed datasets generated during the present study are available from the corresponding author on reasonable request.
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10528_2021_10157_MOESM1_ESM.tif
Supplementary file1 The miR-17-3p was upregulated while FOXO4 was downregulated by TGF-β1. A–D The qRT-PCR assay for the miR-17-3p level analysis (A and B) and western blot for FOXO4 expression detection (C and D) in HaCaT and NHEK cells treated with control (0 ng/mL), 2.5 ng/mL, 5 ng/mL or 10 ng/mL TGF-β1 for 24 h. *P< 0.05 (TIF 743 kb)
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He, Z., Xu, X. Circ_0084443 Inhibits Wound Healing Via Repressing Keratinocyte Migration Through Targeting the miR-17-3p/FOXO4 Axis. Biochem Genet 60, 1236–1252 (2022). https://doi.org/10.1007/s10528-021-10157-5
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DOI: https://doi.org/10.1007/s10528-021-10157-5