Identification of the circ_PRKDC/miR‐20a‐3p/RASA1 axis in regulating HaCaT keratinocyte migration

Abstract

Migration of keratinocytes plays a crucial role in the re‐epithelialization phase during wound healing. Circular RNA (circRNA) protein kinase, DNA‐activated, catalytic subunit (circ_PRKDC, hsa_circ_0084443) has been identified as a regulator of keratinocyte migration. However, the molecular basis governing it remains unclear. The levels of circ_PRKDC, microRNA (miR)‐20a‐3p, and RAS p21 protein activator 1 (RASA1) were assessed by quantitative real‐time PCR (qRT‐PCR) or western blot. Subcellular localization, Actinomycin D, and Ribonuclease (RNase) R assays were performed to characterise circ_PRKDC. Cell migration was gauged by transwell and wound‐healing assays. A direct relationship between miR‐20a‐3p and circ_PRKDC or RASA1 was verified by dual‐luciferase reporter and RNA pull‐down assays. Circ_PRKDC expression was reduced in wound skin during wound healing. Circ_PRKDC modulated migration of HaCaT keratinocytes. Mechanistically, circ_PRKDC directly targeted miR‐20a‐3p. The regulation of circ_PRKDC on HaCaT keratinocyte migration was mediated by miR‐20a‐3p. RASA1 was identified as a direct and functional target of miR‐20a‐3p, and miR‐20a‐3p‐mediated inhibition of RASA1 impacted HaCaT keratinocyte migration. Circ_PRKDC acted as a post‐transcriptional modulator of RASA1 expression through miR‐20a‐3p. Moreover, circ_PRKDC modulated migration of HaCaT keratinocytes by RASA1. Our findings demonstrated a novel molecular basis, the miR‐20a‐3p/RASA1 axis, for the regulation of circ_PRKDC on HaCaT keratinocyte migration.

circCDK13-loaded small extracellular vesicles accelerate healing in preclinical diabetic wound models

“…circRNAs have unique advantages in the development of disease diagnosis and treatment strategies due to their stronger stability, tissue and disease specificity. Recently, accumulating studies have found that circRNAs were closely related to skin wound healing 8,[32][33][34] . In this study, we found that the expression of circCDK13 was down-regulated in diabetic wounds, and then confirmed that circCDK13 could promote the proliferation and migration of HDFs and HEKs in vitro.…”