Zhuang Huafeng, Lin Yongjun, Lin Chengye, Zheng Miao, Li Yizhong, Yao Xuedong, Xu Youjia
Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China.
Department of Orthopedics, The Second Affiliated Hospital of FuJian University, QuanZhou, 362000, China.
J Orthop Surg Res. 2025 Apr 7;20(1):346. doi: 10.1186/s13018-025-05634-1.
To analyze changes in the expression of osteoporosis (OP)-related genes across different bone types based on transcriptome sequencing, and to identify the key molecules and mechanisms involved in the progression of OP in order to better understand this process.
Ten pairs of postmenopausal patients with osteoporosis (OP) and non-osteoporotic (non-OP) volunteers were included. Transcriptome sequencing was performed on six pairs of spongy and cortical bone tissues. The expression of FOXP1 was detected using quantitative real-time PCR (RT-qPCR) and receiver operating characteristic (ROC) curves. Magnetic-activated cell sorting was conducted, and the expression levels of AC003090.1, miR-203a-3p, and FOXP1 were measured using RT-qPCR. Human bone marrow stem cells (hBMSCs) were infected with a lentivirus carrying the AC003090.1 expression plasmid. The expression levels of Runx2, Opn, and Ocn in spongy and cortical bone samples, as well as in post-infection cells, were assessed through RT-qPCR. The expression levels of GSK-3β, β-catenin, and c-Myc were evaluated by performing RT-qPCR and Western blot analysis.
A total of 2,102 out of 2,827 differentially expressed genes (DEGs) were identified between the cortical bone samples from patients with osteoporosis (OP) and the cortical/spongy bone samples of the control group. Among these, 1,482 were significantly up-regulated, and 620 were significantly down-regulated, while 1,146 were significantly up-regulated and 1,681 were significantly down-regulated. The expression of FOXP1 in tissue and bone tissue-derived mesenchymal stem cells (MSCs) from patients with OP was significantly lower than that in patients without OP. FOXP1 levels in bone tissue (cortical bone AUC = 0.825, P = 0.01405; spongy bone AUC = 0.800, P = 0.02338) could serve as predictors of OP. In addition, the overexpression of AC003090.1 significantly enhanced the transcription levels of Runx2, Opn, and Ocn; significantly upregulated the expression levels of β-catenin and c-Myc; and inhibited the expression of GSK-3β. Transfection with miR-203a-3p mimics and FOXP1 small interfering RNA reversed the effect of AC003090.1 on GSK-3β/β-catenin/c-Myc signaling.
FOXP1, as a molecular mediator of AC003090.1, affects the GSK-3β/β-catenin/c-Myc signaling pathway and promotes the osteogenic differentiation of hBMSCs, thus playing a key role in the progression of OP.
基于转录组测序分析不同骨类型中骨质疏松（OP）相关基因的表达变化，鉴定参与OP进展的关键分子和机制，以更好地理解这一过程。
纳入10对绝经后骨质疏松（OP）患者和非骨质疏松（非OP）志愿者。对6对松质骨和皮质骨组织进行转录组测序。采用定量实时PCR（RT-qPCR）和受试者工作特征（ROC）曲线检测FOXP1的表达。进行磁珠分选，并使用RT-qPCR测量AC003090.1、miR-203a-3p和FOXP1的表达水平。用携带AC003090.1表达质粒的慢病毒感染人骨髓干细胞（hBMSCs）。通过RT-qPCR评估松质骨和皮质骨样本以及感染后细胞中Runx2、Opn和Ocn的表达水平。通过RT-qPCR和蛋白质免疫印迹分析评估GSK-3β、β-连环蛋白和c-Myc的表达水平。
在骨质疏松（OP）患者的皮质骨样本与对照组的皮质/松质骨样本之间，共鉴定出2827个差异表达基因（DEG）中的2102个。其中，1482个显著上调，620个显著下调，而在松质骨样本中1146个显著上调，1681个显著下调。OP患者组织和骨组织来源的间充质干细胞（MSC）中FOXP1的表达明显低于非OP患者。骨组织中FOXP1水平（皮质骨AUC = 0.825，P = 0.01405；松质骨AUC = 0.800，P = 0.02338）可作为OP的预测指标。此外，AC003090.1的过表达显著提高了Runx2、Opn和Ocn的转录水平；显著上调了β-连环蛋白和c-Myc的表达水平；并抑制了GSK-3β的表达。用miR-203a-3p模拟物和FOXP1小干扰RNA转染可逆转AC003090.1对GSK-3β/β-连环蛋白/c-Myc信号通路的影响。
FOXP1作为AC003090.1的分子介质，影响GSK-3β/β-连环蛋白/c-Myc信号通路并促进hBMSC的成骨分化，从而在OP进展中起关键作用。
