Tenogenic differentiation of human tendon-derived stem cells induced by long non-coding RNA LINCMD1 via miR-342-3p/EGR1 axis.

IF 2.8 4区医学 Q3 CELL BIOLOGY

Connective Tissue Research Pub Date : 2023-09-01 DOI:10.1080/03008207.2023.2217258

Feng Qu, Xuezhen Shen, Ketao Wang, Chengyi Sun, Pengfei Li

{"title":"Tenogenic differentiation of human tendon-derived stem cells induced by long non-coding RNA LINCMD1 via miR-342-3p/EGR1 axis.","authors":"Feng Qu, Xuezhen Shen, Ketao Wang, Chengyi Sun, Pengfei Li","doi":"10.1080/03008207.2023.2217258","DOIUrl":null,"url":null,"abstract":"Background: Tendon-derived stem cells (TDSCs) are proposed as a potential cell-seed for the treatment of tendon injury due to their tenogenic differentiation potential. In this work, we defined the action of long non-coding RNA (lncRNA) muscle differentiation 1 (LINCMD1) in tenogenic differentiation of human TDSCs (hTDSCs).Methods: Quantitative real-time PCR (qRT-PCR) was used to assess the levels of LINCMD1, microRNA (miR)-342-3p, and early growth response-1 (EGR1) mRNA. Cell proliferation was detected by the XTT colorimetric assay. Protein expression was quantified by western blot. hTDSCs were grown in an osteogenic medium to induce osteogenic differentiation, and the extent of osteogenic differentiation was assessed by Alizarin Red Staining (ARS). The activity of alkaline phosphatase (ALP) was measured by the ALP Activity Assay Kit. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to evaluate the direct relationship between miR-342-3p and LINCMD1 or EGR1.Results: Our results showed that enforced expression of LINCMD1 or suppression of miR-342-3p accelerated the proliferation and tenogenic differentiation and reduced osteogenic differentiation of hTDSCs. LINCMD1 regulated miR-342-3p expression by binding to miR-342-3p. EGR1 was identified as a direct and functional target of miR-342-3p, and knockdown of EGR1 reversed the effects of miR-342-3p suppression on cell proliferation and tenogenic and osteogenic differentiation. Furthermore, the miR-342-3p/EGR1 axis mediated the regulation of LINCMD1 on hTDSC proliferation and tenogenic and osteogenic differentiation.Conclusion: Our study suggests the induction of LINCMD1 in tenogenic differentiation of hTDSCs through miR-342-3p/EGR1 axis.","PeriodicalId":10661,"journal":{"name":"Connective Tissue Research","volume":"64 5","pages":"479-490"},"PeriodicalIF":2.8000,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Connective Tissue Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/03008207.2023.2217258","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CELL BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Background: Tendon-derived stem cells (TDSCs) are proposed as a potential cell-seed for the treatment of tendon injury due to their tenogenic differentiation potential. In this work, we defined the action of long non-coding RNA (lncRNA) muscle differentiation 1 (LINCMD1) in tenogenic differentiation of human TDSCs (hTDSCs).

Methods: Quantitative real-time PCR (qRT-PCR) was used to assess the levels of LINCMD1, microRNA (miR)-342-3p, and early growth response-1 (EGR1) mRNA. Cell proliferation was detected by the XTT colorimetric assay. Protein expression was quantified by western blot. hTDSCs were grown in an osteogenic medium to induce osteogenic differentiation, and the extent of osteogenic differentiation was assessed by Alizarin Red Staining (ARS). The activity of alkaline phosphatase (ALP) was measured by the ALP Activity Assay Kit. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to evaluate the direct relationship between miR-342-3p and LINCMD1 or EGR1.

Results: Our results showed that enforced expression of LINCMD1 or suppression of miR-342-3p accelerated the proliferation and tenogenic differentiation and reduced osteogenic differentiation of hTDSCs. LINCMD1 regulated miR-342-3p expression by binding to miR-342-3p. EGR1 was identified as a direct and functional target of miR-342-3p, and knockdown of EGR1 reversed the effects of miR-342-3p suppression on cell proliferation and tenogenic and osteogenic differentiation. Furthermore, the miR-342-3p/EGR1 axis mediated the regulation of LINCMD1 on hTDSC proliferation and tenogenic and osteogenic differentiation.

Conclusion: Our study suggests the induction of LINCMD1 in tenogenic differentiation of hTDSCs through miR-342-3p/EGR1 axis.

查看原文本刊更多论文

长链非编码RNA LINCMD1通过miR-342-3p/EGR1轴诱导人肌腱源性干细胞的肌腱分化

背景:肌腱源性干细胞(tdsc)因其具有肌腱分化潜能而被认为是治疗肌腱损伤的潜在细胞种子。在这项工作中，我们定义了长链非编码RNA (lncRNA)肌肉分化1 (LINCMD1)在人tdsc (htdsc)的肌腱分化中的作用。方法:采用实时荧光定量PCR (Quantitative real-time PCR, qRT-PCR)检测小鼠外周血LINCMD1、microRNA (miR)-342-3p、早期生长反应-1 (early growth response-1, EGR1) mRNA表达水平。用XTT比色法检测细胞增殖。western blot检测蛋白表达。htdsc在成骨培养基中培养，诱导成骨分化，茜素红染色(ARS)评估成骨分化程度。采用碱性磷酸酶活性测定试剂盒测定碱性磷酸酶(ALP)活性。采用双荧光素酶报告基因和RNA免疫沉淀(RIP)检测来评估miR-342-3p与LINCMD1或EGR1之间的直接关系。结果:我们的研究结果表明，强化表达LINCMD1或抑制miR-342-3p加速htdsc的增殖和成骨分化，降低成骨分化。LINCMD1通过结合miR-342-3p调节miR-342-3p的表达。EGR1被认为是miR-342-3p的直接和功能性靶点，EGR1的敲低逆转了miR-342-3p抑制对细胞增殖和成骨分化的影响。此外，miR-342-3p/EGR1轴介导了LINCMD1对hTDSC增殖和成骨质分化的调节。结论:我们的研究提示LINCMD1通过miR-342-3p/EGR1轴诱导htdsc的成腱分化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Connective Tissue Research 生物-细胞生物学

CiteScore

6.60

自引率

3.40%

发文量

审稿时长

2 months

期刊介绍： The aim of Connective Tissue Research is to present original and significant research in all basic areas of connective tissue and matrix biology. The journal also provides topical reviews and, on occasion, the proceedings of conferences in areas of special interest at which original work is presented. The journal supports an interdisciplinary approach; we present a variety of perspectives from different disciplines, including Biochemistry Cell and Molecular Biology Immunology Structural Biology Biophysics Biomechanics Regenerative Medicine The interests of the Editorial Board are to understand, mechanistically, the structure-function relationships in connective tissue extracellular matrix, and its associated cells, through interpretation of sophisticated experimentation using state-of-the-art technologies that include molecular genetics, imaging, immunology, biomechanics and tissue engineering.