Huang Cong, Geng Junnan, Wei Xiajie, Zhang Ruirui, Jiang Siwen
Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, China.
The Cooperative Innovation Center for Sustainable Pig Production, Wuhan, China.
FEBS Lett. 2016 Mar;590(6):795-807. doi: 10.1002/1873-3468.12112. Epub 2016 Mar 8.
Despite extensive research on osteoblast differentiation and proliferation in mesenchymal stem cells (MSCs), the accurate mechanism remains to be further elucidated. MicroRNAs have been reported to be key regulators of osteoblast differentiation and proliferation. Here, we found that miR-144-3p is down-regulated during osteoblast differentiation of C3H10T1/2 cells. Overexpression of miR-144-3p inhibited osteogenic differentiation, whereas inhibition of miR-144-3p reversed this process. Furthermore, miR-144-3p inhibited the proliferation of C3H10T1/2 cells by arresting cells at the G0/G1 phase. Results from bioinformatics analysis, luciferase assay and western blotting demonstrated that miR-144-3p directly targeted Smad4. Additionally, Smad4 knockdown blocks the effects of miR-144-3p inhibitor. Therefore, we conclude that miR-144-3p negatively regulates osteogenic differentiation and proliferation of C3H10T1/2 cells by targeting Smad4.
尽管对间充质干细胞（MSCs）中破骨细胞的分化和增殖进行了广泛研究，但其精确机制仍有待进一步阐明。据报道，微小RNA是破骨细胞分化和增殖的关键调节因子。在此，我们发现miR-144-3p在C3H10T1/2细胞向破骨细胞分化的过程中表达下调。过表达miR-144-3p可抑制成骨分化，而抑制miR-144-3p则可逆转这一过程。此外，miR-144-3p通过将细胞阻滞在G0/G1期来抑制C3H10T1/2细胞的增殖。生物信息学分析、荧光素酶测定和蛋白质印迹结果表明，miR-144-3p直接靶向Smad4。此外，敲低Smad4可阻断miR-144-3p抑制剂的作用。因此，我们得出结论，miR-144-3p通过靶向Smad4对C3H10T1/2细胞的成骨分化和增殖起负向调节作用。
