miR-452-3p inhibited osteoblast differentiation by targeting Smad4.
miR-452-3p inhibited osteoblast differentiation by targeting Smad4.
复制标题
miR-452-3p 通过靶向 Smad4 抑制成骨细胞分化
作者：
Wu M;Wang H;Kong D;Shao J;Song C;Yang T;Zhang Y
成骨细胞分化是一个复杂的过程，是正常骨形成所必需的。越来越多的研究表明，microRNAs（miRNAs）是包括骨生成在内的多种生理和病理过程的关键调控因子。本研究采用BMP-2诱导MC 3 T3-E1细胞，构建成骨细胞分化模型。通过RT-PCR分析、Western blot分析、ALP活性测定、茜素红染色等方法，研究miR-452- 3 p对成骨细胞分化的影响及其分子机制。我们发现miR-452- 3 p在成骨细胞分化中显著下调。过表达miR-452- 3 p（miR-452- 3 p模拟物）显著抑制成骨细胞标志基因RUNX 2、骨桥蛋白（OPN）和胶原1型a1链（Col 1A 1）的表达，并降低钙结节数量和ALP活性。相反，敲低miR-452- 3 p（miR-452- 3 p抑制剂）产生相反的效果。在机制方面，我们发现Smad 4可能是miR-452- 3 p的靶点，而Smad 4的敲除（si-Smad 4）部分抑制了miR-452- 3 p促进的成骨细胞分化。我们的研究结果表明，miR-452- 3 p通过靶向Smad 4在成骨细胞分化中起重要作用。因此，miR-452- 3 p有望用于骨形成和再生的治疗。
Osteoblast differentiation is a complex process that is essential for normal bone formation. A growing number of studies have shown that microRNAs (miRNAs) are key regulators in a variety of physiological and pathological processes, including osteogenesis. In this study, BMP2 was used to induce MC3T3-E1 cells to construct osteoblast differentiation cell model. Then, we investigated the effect of miR-452-3p on osteoblast differentiation and the related molecular mechanism by RT-PCR analysis, Western blot analysis, ALP activity, and Alizarin Red Staining. We found that miR-452-3p was significantly downregulated in osteoblast differentiation. Overexpression miR-452-3p (miR-452-3p mimic) significantly inhibited the expression of osteoblast marker genes RUNX2, osteopontin (OPN), and collagen type 1 a1 chain (Col1A1), and decreased the number of calcium nodules and ALP activity. In contrast, knockdown miR-452-3p (miR-452-3p inhibitor) produced the opposite effect. In terms of mechanism, we found that Smad4 may be the target of miR-452-3p, and knockdown Smad4 (si-Smad4) partially inhibited the osteoblast differentiation enhanced by miR-452-3p. Our results suggested that miR-452-3p plays an important role in osteoblast differentiation by targeting Smad4. Therefore, miR-452-3p is expected to be used in the treatment of bone formation and regeneration.
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