N(6)-Methyladenosine modification (m(6)A) of circRNA-ZNF638 contributes to the induced activation of SHF stem cells through miR-361-5p/Wnt5a axis in cashmere goats

OBJECTIVE: The objective of this study was to investigate the effects of N(6)-Methyladenosine modification-circRNA-zinc finger protein 638 (m(6)A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats. METHODS: The m(6)A modification of ZNF638 was analyzed using methylation immunoprecipitation with real-time quantitative polymerase chain reaction technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in m(6)A dependence were evaluated through the overexpression of circRNA-ZNF638/its m(6)A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3′-untranslated region was analyzed through Dual-luciferase reporter assay. RESULTS: The m(6)A-circRNA-ZNF638 had significantly higher transcription at anagen SHF bulge of cashmere goats compared with that at telogen, as well as it positively regulated the induced activation of SHF-stem cells in cashmere goats. Mechanismly, m(6)A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal m(6)A modification within circRNA-ZNF638 is required for mediating the miR-361-5p/Wnt5a pathway to regulate the induced activation of SHF stem cells through an introducing of m(6)A-deficient mutant of circRNA-ZNF638. CONCLUSION: The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in m(6)A-dependent manner through miR-361-5p/Wnt5a axis.