lncRNA H19促进胎盘间充质干细胞成骨分化的相关研究  

Study on lncRNA H19 promoting osteogenic differentiation of placenta-derived mesenchymal stem cells

作　　者：卫柄邑 谢艳 吴晓龙 邢伟鹏 王娜 宁桃丽 WEI Bingyi;XIE Yan;WU Xiaolong;XING Weipeng;WANG Na;NING Taoli(Henan Luoyang Orthopaedic Hospital(Henan Orthopaedic Hospital),Luoyang 471002,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院),河南省洛阳市471002

出　　处：《组织工程与重建外科》2024年第6期665-671,共7页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：河南省重点研发与推广专项(科技攻关,212102310855);洛阳市科技计划项目(2101035A)。

摘　　要：目的观察lncRNA H19转染人胎盘间充质干细胞后对多个成骨、成血管因子表达的影响,为构建成骨性能良好的骨缺损填充物提供前期研究基础。方法首先进行调控成骨、成血管因子的miRNAs筛选,将目标锁定为miR-29b-3p、miR-29c-3p、miR-137-3p和miR-383-5p。通过进一步的生物信息学检索,lncRNA H19可以同时调控上述四种miRNAs。进而优选其中调控成骨成血管因子较多的miR-137-3p和miR-383-5p为H19的靶标,利用双荧光素酶报告基因实验验证H19与miR-383-5p、miR-137-3p是否具有调控关系。进一步将H19克隆到慢病毒,然后转染到胎盘间充质干细胞,观察干细胞中miR-137-3p和miR-383-5p表达情况,及IBSP、RUNX2、FGF5和BMP2K的表达能力。通过碱性磷酸酶和茜素红染色,观察转染了H19的胎盘间充质干细胞的成骨性能是否得到提高。结果转染lncRNA H19的人胎盘间充质干细胞内miR-137-3p、miR-383-5p表达下调,而RUNX2、IBSP的表达均明显升高(P<0.05),BMP2K和FGF5的表达各组间无明显差异(P>0.05)。胎盘间充质干细胞转染H19后成骨性能得到提升。结论转染lncRNA H19的人胎盘间充质干细胞可以促进某些成骨成血管因子的表达且提高成骨性能,为今后研制成骨性能良好的组织工程骨提供了可能。Objective To observe the effects of lncRNA H19 transfection on the expression of multiple osteogenic and angiogenic factors in human placenta-derived mesenchymal stem cells(P-MSCs),and to provide a preliminary research foundation for the synthesis of bone defect fillers with good osteogenic properties in the future.Methods First,miRNAs that regulate a large number of osteogenic and angiogenic factors were screened,and miR-29b-3p,miR-29c-3p,miR-137-3p,and miR-383-5p were identified as the target miRNAs.Further bioinformatics analysis revealed that lncRNA H19 can regulate all of these miRNAs.Subsequently,miR-137-3p and miR-383-5p,which regulate a greater number of osteogenic and angiogenic factors,were selected as the targets of H19.Dual luciferase reporter gene assay was used to verify the regulatory relationship between H19 and miR-383-5p/miR-137-3p.H19 was cloned into lentivirus and transfected into placenta-derived mesenchymal stem cells(PMSCs).The expression levels of miR-137-3p,miR-383-5p,IBSP,RUNX2,FGF5 and BMP2K were observed.Alkaline phosphatase and Alizarin red staining were used to assess the improved osteogenic potential of H19-transfected P-MSCs.Results Transfection of lncRNA H19 downregulated the expression of miR-137-3p and miR-383-5p in human P-MSCs,and significantly upregulated the expression of RUNX2 and IBSP(P<0.05),and there were no significant differences in BMP2K and FGF5 expression between groups.Furthermore,osteogenic performance was enhanced in P-MSCs transfected with H19.Conclusion Transfection of lncRNA H19 in human placenta-derived mesenchymal stem cells can promote the expression of certain osteogenic and angiogenic factors,and improve the osteogenic performance,which provides a possibility for the development of tissue-engineered bone with good osteogenic properties in the future.

关 键 词：长链非编码RNA H19 胎盘间充质干细胞 成骨成血管因子 

分 类 号：Q813.11[生物学—生物工程]