MicroRNA-125b-5p inhibits proliferation and promotes adipogenic differentiation in 3T3-L1 preadipocytes
MicroRNA-125b-5p inhibits proliferation and promotes adipogenic differentiation in 3T3-L1 preadipocytes
复制标题
MicroRNA-125b-5p 抑制 3T3-L1 前脂肪细胞增殖并促进成脂分化
DOI：
10.1093/abbs/gmv024
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发表时间：
2015-05-01
影响因子：
3.7
通讯作者：
Li, Yugu
中科院分区：
文献类型：
--
作者：
Ouyang, Dan;Ye, Yaqiong;Li, Yugu
先前的证据表明microRNA-125 b（miR-125 b）家族在调节癌细胞生长、发育、分化和凋亡中起重要作用。然而，它们是否有助于脂肪细胞分化的过程仍不清楚。在本研究中，我们发现miR-125 b家族成员miR-125 b-5 p的表达水平在3 T3-L1前脂肪细胞向成熟脂肪细胞分化的过程中显著上调。向3 T3-L1细胞中补充miR-125 b-5 p可促进脂肪形成分化，这可通过脂滴和脂肪细胞特异性分子标记物（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）的mRNA水平增加以及甘油三酯蓄积来证明。CCK-8法显示miR-125 b-5 p补充显著抑制细胞增殖。流式细胞术分析显示，miR-125 b-5 p损害G1/S期转换以及G1/S相关基因的mRNA和蛋白表达，如Cyclin D2、Cyclin D3和CDK 4。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smad 4可能是小鼠3 T3-L1前脂肪细胞中miR-125 b-5 p的潜在靶点，解释了miR-125 b-5 p的一些功能。总而言之，这些数据表明miR-125 b-5 p可能是脂肪细胞分化的重要正调节因子，至少部分通过下调smad 4来实现。
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 gamma, CCAAT/enhancer-binding protein alpha, 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.