Zhong-Zheng Zhi, Tao Liu, Jian Kang, Fu-Chao Zhou, Xiao-Dong Liu, Zhi-Min He
{"title":"miR-365通过与HIF-1α相互作用促进hoxa9介导的间充质干细胞向髓核细胞的分化。","authors":"Zhong-Zheng Zhi, Tao Liu, Jian Kang, Fu-Chao Zhou, Xiao-Dong Liu, Zhi-Min He","doi":"10.5312/wjo.v16.i7.107045","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Degenerative disc disease (DDD) is characterized by the loss of nucleus pulposus cells (NPCs). Inducing differentiation of bone marrow mesenchymal stem cells (MSCs) into NPCs has emerged as a novel therapeutic strategy for DDD. However, the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.</p><p><strong>Aim: </strong>To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.</p><p><strong>Methods: </strong><i>In vitro</i>, the effects of miR-365 on MSC proliferation, apoptosis, and differentiation were assessed by cell counting kit-8 assay, flow cytometry, and quantitative real-time polymerase chain reaction (qRT-PCR). <i>In vivo</i>, the expression levels of miR-365, HIF-1α, Sox9, Kdm6a<i>,</i> and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.</p><p><strong>Results: </strong><i>In vitro</i>, miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis. The expression levels of glycosaminoglycans, proteoglycan, and type 2 collagen were significantly increased with miR-365 ectopic expression. <i>In vivo</i>, the expression levels of miR-365, HIF-1α, Sox9, and Kdm6a were significantly increased, whereas HOXA9 was remarkably decreased. Mechanically, miR-365 inhibited HOXA9 expression by directly binding to its 3' untranslated region. HOXA9 could inhibit HIF-1α expression by binding to the <i>Hif-1α</i> promoter, thereby affecting the expression levels of <i>Sox9</i> and <i>Kdm6a</i>. Moreover, HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.</p><p><strong>Conclusion: </strong>miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1α and may serve as a potential target for DDD treatment.</p>","PeriodicalId":47843,"journal":{"name":"World Journal of Orthopedics","volume":"16 7","pages":"107045"},"PeriodicalIF":2.3000,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12278291/pdf/","citationCount":"0","resultStr":"{\"title\":\"miR-365 promotes HOXA9-mediated differentiation of mesenchymal stem cells to nucleus pulposus cells by interacting with HIF-1α.\",\"authors\":\"Zhong-Zheng Zhi, Tao Liu, Jian Kang, Fu-Chao Zhou, Xiao-Dong Liu, Zhi-Min He\",\"doi\":\"10.5312/wjo.v16.i7.107045\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Degenerative disc disease (DDD) is characterized by the loss of nucleus pulposus cells (NPCs). Inducing differentiation of bone marrow mesenchymal stem cells (MSCs) into NPCs has emerged as a novel therapeutic strategy for DDD. However, the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.</p><p><strong>Aim: </strong>To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.</p><p><strong>Methods: </strong><i>In vitro</i>, the effects of miR-365 on MSC proliferation, apoptosis, and differentiation were assessed by cell counting kit-8 assay, flow cytometry, and quantitative real-time polymerase chain reaction (qRT-PCR). <i>In vivo</i>, the expression levels of miR-365, HIF-1α, Sox9, Kdm6a<i>,</i> and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.</p><p><strong>Results: </strong><i>In vitro</i>, miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis. The expression levels of glycosaminoglycans, proteoglycan, and type 2 collagen were significantly increased with miR-365 ectopic expression. <i>In vivo</i>, the expression levels of miR-365, HIF-1α, Sox9, and Kdm6a were significantly increased, whereas HOXA9 was remarkably decreased. Mechanically, miR-365 inhibited HOXA9 expression by directly binding to its 3' untranslated region. HOXA9 could inhibit HIF-1α expression by binding to the <i>Hif-1α</i> promoter, thereby affecting the expression levels of <i>Sox9</i> and <i>Kdm6a</i>. Moreover, HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.</p><p><strong>Conclusion: </strong>miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1α and may serve as a potential target for DDD treatment.</p>\",\"PeriodicalId\":47843,\"journal\":{\"name\":\"World Journal of Orthopedics\",\"volume\":\"16 7\",\"pages\":\"107045\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2025-07-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12278291/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"World Journal of Orthopedics\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.5312/wjo.v16.i7.107045\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ORTHOPEDICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"World Journal of Orthopedics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.5312/wjo.v16.i7.107045","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ORTHOPEDICS","Score":null,"Total":0}
miR-365 promotes HOXA9-mediated differentiation of mesenchymal stem cells to nucleus pulposus cells by interacting with HIF-1α.
Background: Degenerative disc disease (DDD) is characterized by the loss of nucleus pulposus cells (NPCs). Inducing differentiation of bone marrow mesenchymal stem cells (MSCs) into NPCs has emerged as a novel therapeutic strategy for DDD. However, the efficiency of MSC differentiation and the underlying mechanisms remain to be fully elucidated.
Aim: To investigate the role and mechanism of miR-365 in promoting the differentiation of MSCs into NPCs for DDD treatment.
Methods: In vitro, the effects of miR-365 on MSC proliferation, apoptosis, and differentiation were assessed by cell counting kit-8 assay, flow cytometry, and quantitative real-time polymerase chain reaction (qRT-PCR). In vivo, the expression levels of miR-365, HIF-1α, Sox9, Kdm6a, and HOXA9 in the spinal cord of rats with spinal cord injury were determined by qRT-PCR and Western blot.
Results: In vitro, miR-365 significantly promoted MSC proliferation and inhibited MSC apoptosis. The expression levels of glycosaminoglycans, proteoglycan, and type 2 collagen were significantly increased with miR-365 ectopic expression. In vivo, the expression levels of miR-365, HIF-1α, Sox9, and Kdm6a were significantly increased, whereas HOXA9 was remarkably decreased. Mechanically, miR-365 inhibited HOXA9 expression by directly binding to its 3' untranslated region. HOXA9 could inhibit HIF-1α expression by binding to the Hif-1α promoter, thereby affecting the expression levels of Sox9 and Kdm6a. Moreover, HOXA9 knockdown significantly reversed the differentiation of MSCs into NPCs induced by miR-365.
Conclusion: miR-365 promotes HOXA9-mediated differentiation of MSCs into NPCs by interacting with HIF-1α and may serve as a potential target for DDD treatment.