Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur 603203, Tamil Nadu, India.
Department of Endocrinology, Dr. A.L.M. PG Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai 600 113, Tamil Nadu, India.
Int J Biol Macromol. 2019 Jul 1;132:541-549. doi: 10.1016/j.ijbiomac.2019.04.003. Epub 2019 Apr 3.
Transforming growth factor-beta1 (TGF-β1) is a pleiotropic and ubiquitous cytokine involved in bone development and bone remodeling. Matrix metalloproteinase-13 (MMP13) plays a role in the degradation of the extracellular matrix (ECM), and the regulation of this gene is critical in bone remodeling. We previously reported that TGF-β1 stimulates MMP13 expression in rat osteoblasts. Recently, studies have examined the regulation of bone metabolism by microRNAs (miRNAs) to determine their therapeutic potential in osteogenesis. Here, we assessed the effect of TGF-β1 on down-regulation of miRNAs that target MMP13 and stimulation of MMP13 expression in osteoblasts. We used in silico analysis and identified 11 specific miRNAs which directly target rat MMP13. Among these miRNAs, miR-203a-5p expression was significantly decreased by TGF-β1-treatment in rat osteoblasts. Transient transfection of a miR-203a-5p mimic into rat osteoblasts reduced MMP13 expression. A luciferase reporter assay confirmed a direct targeting of miR-miR-203a-5p with the 3' untranslated regions of the MMP13 gene. Hence, we suggest that TGF-β1 stimulated down-regulation of miR-203a-5p, resulting in the stimulation of MMP13 expression in rat osteoblasts. Thus, identification of the role of miR-203a-5p via TGF-β1 and MMP13 in bone remodeling indicated its potential as a biomarker or therapeutic agent for treating bone and bone-related diseases.
转化生长因子-β1（TGF-β1）是一种多功能且普遍存在的细胞因子，参与骨发育和骨重塑。基质金属蛋白酶-13（MMP13）在细胞外基质（ECM）的降解中起作用，该基因的调节在骨重塑中至关重要。我们之前报道过 TGF-β1 刺激大鼠成骨细胞中 MMP13 的表达。最近，研究已经检查了 microRNAs（miRNAs）对骨代谢的调节，以确定它们在成骨中的治疗潜力。在这里，我们评估了 TGF-β1 对针对 MMP13 的 miRNAs 的下调以及对成骨细胞中 MMP13 表达的刺激作用。我们使用计算机分析并鉴定了 11 种直接靶向大鼠 MMP13 的特定 miRNAs。在这些 miRNAs 中，miR-203a-5p 的表达在 TGF-β1 处理大鼠成骨细胞后显著降低。瞬时转染 miR-203a-5p 模拟物可降低大鼠成骨细胞中 MMP13 的表达。荧光素酶报告基因分析证实了 miR-203a-5p 与 MMP13 基因 3'非翻译区的直接靶向作用。因此，我们认为 TGF-β1 刺激 miR-203a-5p 的下调，导致大鼠成骨细胞中 MMP13 的表达受到刺激。因此，通过 TGF-β1 和 MMP13 鉴定 miR-203a-5p 在骨重塑中的作用表明其作为治疗骨和骨相关疾病的生物标志物或治疗剂的潜力。
