LncRNA MTC enhances the proliferation of Liaoning cashmere goat skin fibroblasts by modulating GSTM1-ASK1 interaction

抄録

Cashmere quality represents a crucial economic trait in cashmere goats. Investigating methods to enhance cashmere yield and quality is essential for the sustainable development of the cashmere industry. Although melatonin (MT) has been shown to promote cashmere wool growth, the underlying molecular mechanisms, particularly those involving melatonin-mediated long non-coding RNA (lncRNA), remain poorly understood. In this study, cashmere growth-related lncRNA (LncRNA MTC) was used as the starting point. LncRNA MTC overexpression and interference lentivirus vectors were constructed and transfected into Liaoning cashmere goat skin fibroblasts. Results from the cholecystokinin-octapeptide (CCK-8) assay demonstrated that LncRNA MTC overexpression significantly enhanced cell proliferation, while LncRNA MTC interference notably suppressed it. RNA pull-down coupled with mass spectrometry revealed a significant increase in LncRNA MTC’s binding to glutathione S-transferase M1 (GSTM1) in Liaoning cashmere goat skin fibroblasts following melatonin treatment. RNA immunoprecipitation (RIP) technology confirmed that the intracellular GSTM1 protein indeed interacts with LncRNA MTC. CCK-8 results showed that interference with GSTM1 could reverse the promotion of MT on the proliferation of skin fibroblasts of Liaoning cashmere goats. Co-immunoprecipitation (Co-IP) results demonstrated an association between GSTM1 and apoptosis signal-regulating kinase 1 (ASK1) proteins in Liaoning cashmere goat skin fibroblasts. The findings indicate that melatonin-mediated LncRNA MTC, which is related to cashmere growth, binds to the GSTM1 protein. This interaction affects the GSTM1-ASK1 protein complex formation, consequently inhibiting the apoptosis of Liaoning cashmere goat skin fibroblasts.