RRM2 regulates osteogenesis of mouse embryo fibroblasts via the Wnt/beta-catenin signaling pathway

作者全名："Cai, Haijun; Guo, Hui; Deng, Yixuan; Jiang, Jinhai; Liu, Siyuan; He, Wenge; Jian, Huagang"

作者地址："[Cai, Haijun; Guo, Hui; Jian, Huagang] Chongqing Med Univ, Affiliated Hosp 2, Dept Emergency, Chongqing 400010, Peoples R China; [Deng, Yixuan; Jiang, Jinhai] Chongqing Med Univ, Sch Pharm, Dept Pharmacol, Chongqing 400016, Peoples R China; [Liu, Siyuan; He, Wenge] Chongqing Med Univ, Affiliated Hosp 2, Dept Orthoped, Chongqing 400010, Peoples R China; [Jian, Huagang] Chongqing Med Univ, Dept Emergency, Affiliated Hosp 2, 74 Linjiang Rd, Chongqing 400010, Peoples R China"

通信作者："Jian, HG (通讯作者)，Chongqing Med Univ, Dept Emergency, Affiliated Hosp 2, 74 Linjiang Rd, Chongqing 400010, Peoples R China."

来源：EXPERIMENTAL AND THERAPEUTIC MEDICINE

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:000890912800001

JCR分区：Q3

影响因子：2.7

年份：2022

卷号：24

期号：4

开始页：　

结束页：　

文献类型：Article

关键词：ribonucleotide reductase M2; osteoporosis; mouse embryo fibroblasts; osteogenic differentiation; Wnt/beta-catenin

摘要："Osteoporosis is a widespread bone metabolic disease characterized by reduced bone mass and bone microstructure deterioration. Ribonucleotide reductase M2 (RRM2) is a key enzyme in DNA synthesis and repair. The present study investigated the effect of RRM2 on osteogenesis of mouse embryo fibroblasts (MEFs) and its molecular mechanism. Bioinformatics analysis revealed that RRM2 expression was increased during osteogenesis of MEFs triggered by bone morphogenetic protein 9. Subsequently, MEFs were used as a mesenchymal stem cell model and osteogenic inducing medium was used to induce osteogenic differentiation. RRM2 protein expression was measured by western blotting during osteogenic differentiation induction of MEFs. RRM2 levels in MEFs were upregulated and downregulated by RRM2-overexpressing recombinant adenovirus and small interfering RNA-RRM2, respectively. Bone formation markers (RUNX family transcription factor 2, osterix, distal-less homeobox 5, collagen type I alpha 1 chain, osteopontin and osteocalcin) were detected by reverse transcription-quantitative (RT-q) PCR and alkaline phosphatase (ALP) and Alizarin Red S staining were examined. The protein expression levels of beta-catenin and the ratio of phosphorylated (p-)GSK-3 beta to GSK-3 beta were detected by western blotting and the RNA expression of downstream related target genes (beta-catenin, axis inhibition protein 2 (AXIN2), transcription factor 7 like 2, lymphoid enhancer binding factor 1, c-MYC and Cyclin D1) in the Wnt/beta-catenin signaling pathway was measured by RT-qPCR. RRM2 protein expression increased as the osteogenic differentiation induction period was extended. RRM2 overexpression increased osteogenic marker RNA expression, ALP activity, bone mineralization, the protein expression levels of beta-catenin, the ratio of p-GSK-3 beta to GSK-3 beta and the RNA expression of downstream related target genes in the Wnt/beta-catenin signaling pathway, whereas RRM2 knockdown had the opposite effect. The findings of the present study revealed that RRM2 overexpression enhanced osteogenic differentiation, while RRM2 knockdown reduced osteogenic differentiation. RRM2 may regulate osteogenic differentiation of MEFs via the canonical Wnt/beta-catenin signaling pathway, providing a possible therapeutic target for osteoporosis."

基金机构：Trauma Repair Specialty Alliance; [2019jstg025]

基金资助正文："Trauma Repair Specialty Alliance (established on December 29, 2017); Standardized treatment of chronic defect wounds, grant no. 2019jstg025 (to HJ)."