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Existing experiments have found a new intergenic lncRNA activated by melatonin, which is called lncRNA MTC. However, the regulatory mechanism of lncRNA MTC in Liaoning Cashmere goat skin fibroblasts has not been clarified. Specific knockdown of lncRNA MTC inhibits cell proliferation and increases apoptosis. iTRAQ reagent was used for relative and absolute quantification of proteins in lncRNA MTC-KD and NC groups to evaluate changes in protein expression during dermal fibroblast development following lncRNA MTC deletion. A total of 5931 proteins were found in Liaoning Cashmere goat skin fibroblasts, of which 123 were differentially expressed, including 32 up-regulated proteins and 91 down-regulated proteins. Of the 91 down-regulated proteins, 32 act mainly through related pathways (e.g., cell cycle, mitochondrial function, ribosomal structure, vesicular transport, cytoskeletal components and skin morphogenesis). LncRNA MTC facilitates the proliferation of Liaoning Cashmere goat skin fibroblasts by regulating ITGB5, TlN2, CTSS, POLG, RAP1B, CHAF1A, CDCA8 and other proteins involved in cell proliferation. The results of this study provide some candidate proteins for the in-depth investigation of the molecular mechanism of lncRNA MTC, which facilitates hair growth in cashmere goats and provides more insights into their regulatory networks and biochemical pathways. [ABSTRACT FROM AUTHOR]

The Liaoning cashmere goat is a unique breed in China. It has an excellent germplasm resource with high villus yield, fine villus fibre quality, stable genetic performance and significant economic benefits.In this study, the effects of different concentrations of 1.25(OH)2D3 on the proliferation of skin fibroblasts was detected via CCK8 assay, while the expression of genes and proteins related to villus growth in Liaoning cashmere goat were detected by qRT-PCR and Western Blot. Treatment of Liaoning cashmere goat skin fibroblasts with 0.01 ng/L of 1.25(OH)2D3 for 72 h significantly promoted the proliferation of Liaoning cashmere goat skin fibroblasts, increased the expression of VDR, WNT10B, LEF1 and β-catenin genes, and decreased the expression of BMP4 and BMPRIB genes, indicating that 1.25(OH)2D3 plays a positive role in regulating hair follicle growth and development through the WNT and BMP signaling pathway, which positively regulates the growth and development of hair follicles. VDR interacting proteins were screened using Co-IP combined mass spectrometry. After treatment of cells with 0.01 ng/L of 1.25(OH)2D3 for 72 h, VDR interacting proteins were enriched to PI3K-AKT signaling pathways associated with hair follicle growth and development. Moreover, mitogen-activated protein kinase, protein S100, protein disulfide bond isomerase protein, FLII actin remodeling protein and multifunctional proteoglycan protein related to the growth and development of hair follicles were upregulated, indicating that a positive regulatory effect of 1.25(OH)2D3 on hair follicle growth and development. [ABSTRACT FROM AUTHOR]

In this study, the genes related to the Downy growth of Liaoning cashmere goats were screened for their expression with simultaneous melatonin administration, so as to investigate the effects of target genes on the proliferation of skin fibroblasts in this animal species. Genes related to the villus growth of skin fibroblasts were screened by in vitro transcriptome sequencing and verified by qPCR. In addition, gene overexpression and interference were used to study the effects of target genes on the proliferation of skin fibroblasts. Groups treated with M1_24H, M2_24H and M2_72H exhibited significant differences compared with the control group. Among them, the differentially expressed transcripts in the M2_72H group were significantly enriched in the TNF and NOD-like receptor signaling pathways, which are associated with the villus. In addition, eight differentially expressed genes were screened from the TNF and the NOD-like receptor signaling pathways. Verification by qPCR showed that the expression of TNF-α, IL-6, TNFAIP3, PYCARD and NFKBIA genes were significantly upregulated, which was consistent with the sequencing results. Melatonin treatments can significantly lead to an increase in the expression of IL-6 and TNF-α genes. Besides, melatonin treatments can affect cashmere growth in Liaoning cashmere goats by regulating several signaling pathways, including TNF, NOD-like receptor and NF-κB. [ABSTRACT FROM AUTHOR]

The relationship between PLP2 gene and cashmere fiber quality of Liaoning cashmere goat was investigated. The sheep fibroblast cells were treated with exogenous cytokines and melatonin, independently, and RNA interference, RT-PCR and in situ hybridization were utilized for investigating the PLP2 gene regulation mechanism underlying the Liaoning cashmere growth. The results showed that the expression of PLP2 gene in the prosperous and degenerative stage is higher than that of the primary follicle, indicating that the PLP2 gene promotes the secondary follicle, wherein the gene is expressed only in the inner root sheath, suggesting its correlation to hair loss. The results of RT-PCR showed that the trend of FGF5 expression in PLP2 gene was positively regulated. The influence of MT on the expression of PLP2 gene was negatively regulated, and the inhibition was gradually enhanced with the passage of time. Studies have confirmed that the Noggin gene is an inhibitor of the BMP signaling pathway. After the noggin gene interferes with the lentivirus infection, the expression of the PLP2 gene is downregulated. Therefore, the PLP2 gene, along with the other suppressor genes including the noggin gene, might affect the development of hair follicles by inhibiting the BMP（Bone morphogenetic proteins）pathway. [ABSTRACT FROM AUTHOR]