School of Life Sciences, Liaoning Key Laboratory of Biotechnology and Molecular Drug Development, Liaoning Normal University, Dalian, 116081, China.
Mol Biol Rep. 2024 Aug 19;51(1):920. doi: 10.1007/s11033-024-09851-5.
Liaoning cashmere goat is recognized as a valuable genetic resource breed, with restrictions on genetic outflow in China. Hair follicle development in the cashmere goat is influenced by melatonin and long non-coding RNAs (lncRNAs). However, the role of lncRNAs in facilitating melatonin-promoted cashmere growth remains poorly understood. Previous studies have identified a new lncRNA, lncRNA018392, which is involved in the melatonin-promoted proliferation of cashmere skin fibroblasts.
Flow cytometry and CCK-8 assays confirmed that silencing lncRNA018392 negates the effects of melatonin on cell proliferation, and that proliferation was reduced when the gene CSF1R, located near lncRNA018392, was inhibited. Further investigation using a dual-luciferase reporter assay showed that lncRNA018392 could positively regulate the promoter of CSF1R.
Results from RNA-binding protein immunoprecipitation (RIP) and chromatin immunoprecipitation sequencing (ChIP-Seq) revealed that lncRNA018392 interacts with the transcription factor SPI1, with CSF1R being a downstream target gene regulated by SPI1. This interaction was confirmed by ChIP-PCR, which demonstrated SPI1's binding to CSF1R.
This study found that the melatonin-responsive lncRNA018392 accelerates the cell cycle and promotes cell proliferation by recruiting SPI1 to upregulate the expression of the neighboring gene CSF1R. These findings provide a theoretical foundation for elucidating the molecular mechanisms of cashmere growth and for the molecular breeding of cashmere goats.
辽宁绒山羊被认为是一种有价值的遗传资源品种,在中国对其基因外流有严格限制。褪黑素和长链非编码 RNA(lncRNA)影响绒山羊的毛囊发育。然而,lncRNA 促进褪黑素促进羊绒生长的作用仍知之甚少。先前的研究已经确定了一种新的 lncRNA,lncRNA018392,它参与了褪黑素促进的绒山羊皮肤成纤维细胞增殖。
流式细胞术和 CCK-8 检测证实,沉默 lncRNA018392 可消除褪黑素对细胞增殖的影响,而当位于 lncRNA018392 附近的基因 CSF1R 被抑制时,增殖减少。进一步的双荧光素酶报告基因检测证实,lncRNA018392 可以正向调控 CSF1R 的启动子。
RNA 结合蛋白免疫沉淀(RIP)和染色质免疫沉淀测序(ChIP-Seq)的结果表明,lncRNA018392 与转录因子 SPI1 相互作用,CSF1R 是 SPI1 调节的下游靶基因。ChIP-PCR 证实了 SPI1 与 CSF1R 的结合。
本研究发现,褪黑素反应性 lncRNA018392 通过募集 SPI1 上调邻近基因 CSF1R 的表达,加速细胞周期并促进细胞增殖。这些发现为阐明羊绒生长的分子机制和绒山羊的分子育种提供了理论基础。