Ouyang Dan, Ye Yaqiong, Guo Dongguang, Yu Xiaofang, Chen Jian, Qi Junjie, Tan Xiaotong, Zhang Yuan, Ma Yongjiang, Li Yugu
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
Acta Biochim Biophys Sin (Shanghai). 2015 May;47(5):355-61. doi: 10.1093/abbs/gmv024.
Previous evidence has indicated that the microRNA-125b (miR-125b) family plays important roles in the regulation of cancer cell growth, development, differentiation, and apoptosis. However, whether they contribute to the process of adipocyte differentiation remains unclear. In the present study, we revealed that the expression level of miR-125b-5p, a member of miR-125b family, was dramatically up-regulated during differentiation of 3T3-L1 preadipocyte into mature adipocyte. Supplement of miR-125b-5p into 3T3-L1 cells promoted adipogenic differentiation as evidenced by increased lipid droplets and mRNA levels of adipocyte-specific molecular markers, including peroxisome proliferators-activated receptor γ, CCAAT/enhancer-binding protein α, fatty acid-binding protein 4, and lipoprotein lipase, and by triglyceride accumulation. CCK-8 assay showed that miR-125b-5p supplementation significantly inhibited cell proliferation. Flow cytometry analysis showed that miR-125b-5p impaired G1/S phase transition as well as the mRNA and protein expression of G1/S-related genes, such as Cyclin D2, Cyclin D3, and CDK4. Nevertheless, it had no effect on apoptosis. Additionally, by target gene prediction, we demonstrated that smad4 may be a potential target of miR-125b-5p in mouse 3T3-L1 preadipocytes, accounting for some of miR-125b-5p's functions. Taken together, these data indicated that miR-125b-5p may serve as an important positive regulator in adipocyte differentiation, at least partially through down-regulating smad4.
先前的证据表明,微小RNA - 125b(miR - 125b)家族在癌细胞的生长、发育、分化和凋亡调控中发挥重要作用。然而,它们是否参与脂肪细胞分化过程仍不清楚。在本研究中,我们发现miR - 125b家族成员之一miR - 125b - 5p在3T3 - L1前脂肪细胞分化为成熟脂肪细胞的过程中表达水平显著上调。向3T3 - L1细胞中补充miR - 125b - 5p可促进脂肪生成分化,这可通过脂滴增加以及脂肪细胞特异性分子标志物的mRNA水平升高得以证明,这些标志物包括过氧化物酶体增殖物激活受体γ、CCAAT/增强子结合蛋白α、脂肪酸结合蛋白4和脂蛋白脂肪酶,同时甘油三酯也有积累。CCK - 8检测表明,补充miR - 125b - 5p可显著抑制细胞增殖。流式细胞术分析表明,miR - 125b - 5p损害G1/S期转换以及G1/S相关基因如细胞周期蛋白D2、细胞周期蛋白D3和细胞周期蛋白依赖性激酶4的mRNA和蛋白表达。然而,它对细胞凋亡没有影响。此外,通过靶基因预测,我们证明smad4可能是小鼠3T3 - L1前脂肪细胞中miR - 125b - 5p的潜在靶标,这解释了miR - 125b - 5p的部分功能。综上所述,这些数据表明miR - 125b - 5p可能至少部分通过下调smad4作为脂肪细胞分化中的重要正调控因子。