人骨髓间充质干细胞成骨分化过程中差异翻译谱分析。

Hua Liu, Zhi Peng Fan, Qiu Bo Yang, Hui Na Liu
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引用次数: 0

摘要

目的:探讨人颌骨骨髓间充质干细胞(h-JBMMSCs)在成骨分化过程中的差异翻译谱和编码产物。方法:采用Ribo-seq检测h-JBMMSCs的差异翻译基因(DEGs)、开放阅读框(ORFs)和成骨分化期相关基因。Western blotting (WB)检测骨钙素(OCN)和骨涎蛋白(BSP)的表达。碱性磷酸酶(ALP)活性和茜素红染色(ARS)检测成骨分化。设计含有5'UTR-ORF-GFPmut的慢病毒转染h-JBMMSCs,分析荧光和绿色荧光蛋白(GFP)的表达。合成SNHG1肽用于成骨诱导和检测成骨标志物。结果:共检测到53,432个orf,去除带注释的蛋白编码基因后,鉴定出包括lncRNA SNHG1在内的199个候选翻译sorf。此外,5'UTR-ORF-GFPmut显示绿色荧光,表达GFP。lncRNA SNHG1的敲低可提高h-JBMMSCs的ALP活性,促进OCN和BSP的表达,增强ARS强度和钙离子含量。然而,lncRNA SNHG1和SNHG1多肽的过表达抑制了h-JBMMSCs的成骨分化。结论:LncRNA SNHG1抑制h-JBMMSCs成骨分化。LncRNA SNHG1编码一个19个氨基酸的肽,抑制h-JBMMSCs的成骨分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of Differential Translation Profiles of Human Bone Marrow Mesenchymal Stem Cells during Osteogenic Differentiation.

Objective: To explore the differential translation profiles and coding products of human jaw bone marrow mesenchymal stem cells (h-JBMMSCs) during osteogenic differentiation.

Methods: Ribo-seq was used to examine the differential translated genes (DEGs), open reading frames (ORFs) and genes associated with the osteogenic differentiation phase of h-JBMMSCs. Western blotting (WB) was performed to detect the expression of osteocalcin (OCN) and bone sialoprotein (BSP). Alkaline phosphatase (ALP) activity and alizarin red staining (ARS) were used to detect osteogenic differentiation. A lentivirus containing 5'UTR-ORF-GFPmut was designed to transfect h-JBMMSCs, and fluorescence and green fluorescent protein (GFP) expression were analysed. The SNHG1 peptide was synthesised for osteogenic induction and to detect osteogenic markers.

Results: A total of 53,432 ORFs were detected and 199 candidate translation sORFs, including lncRNA SNHG1, were identified after removing the annotated protein-coding genes. In addition, the 5'UTR-ORF-GFPmut showed green fluorescence and expressed GFP. Knockdown of the lncRNA SNHG1 increased the ALP activity of h-JBMMSCs, promoted the expression of OCN and BSP, and enhanced the intensity of ARS and calcium ion content. However, overexpression of lncRNA SNHG1 and the SNHG1 polypeptide inhibited the osteogenic differentiation of h-JBMMSCs.

Conclusion: LncRNA SNHG1 inhibited the osteogenic differentiation of h-JBMMSCs. LncRNA SNHG1 can encode a peptide of 19-amino acid and inhibit the osteogenic differentiation of h-JBMMSCs.

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