miR-497-5p通过调控RSPO3影响绒山羊毛乳头细胞增殖的研究OA

This study aimed to reveal the molecular regulatory mechanism of differentially expressed miR-497-5p in dermal papilla cells during the induction(E65),organogenesis(E90),and cytodifferentiation(E120)stages of cashmere goats,providing a theoretical basis for improving cashmere yield.In this study,we analyzed the miRNA-seq data of embryonic skin tissues of cashmere goats and counted the differential expression of miR-497-5p in the periods of E65,E90,and E120.Potential target genes associated with hair follicle development were predicted using cross-analyzed with RNA-seq.The targeting relationship between miR-497-5p and the potential target gene RSPO3 was verified using a dual luciferase reporter system.The miR-497-5p mimics/mimics nc and inhibitor/inhibitor nc were transfected into dermal papilla cells.The expression of RSPO3 at the mRNA and protein levels was detected by RT-qPCR,WB and immunofluorescence,and the targeting relationship between miR-497-5p and RSPO3 was further verified using functional rescue assays.A total of 252 miR-497-5p-targeted embryonic differential genes were screened,and the results of GO enrichment analysis showed that terms such as regulation of developmental growth were significantly enriched.Dual luciferase assay showed that miR-497-5p could bind to the 3'UTR of RSPO3.RT-qPCR,WB and immunofluorescence showed that the overexpression of miR-497-5p resulted in a significant decrease in the expression of RSPO3 at the mRNA and protein levels(P＜0.05),and RSPO3 expression was significantly increased after miR-497-5p inhibition(P＜0.05).The results of functional rescue assay showed that cell viability was significantly reduced after simultaneous overexpression of miR-497-5p and RSPO3 compared with the RSPO3-only group(P＜0.01).This study indicated that miR-497-5p regulated the proliferation of dermal papilla cells through targeting RSPO3,thereby revealing its molecular mechanism in hair follicle development of cashmere goat.