The Fto Gene Regulates the Proliferation and Differentiation of Pre-Adipocytes in Vitro.
The Fto Gene Regulates the Proliferation and Differentiation of Pre-Adipocytes in Vitro.
复制标题
Fto基因在体外调控前脂肪细胞的增殖和分化
作者：
Jiao Y;Zhang J;Lu L;Xu J;Qin L
高度调控的前脂肪细胞的分化和增殖在肥胖的发生中起着关键作用。脂肪量与肥胖相关基因（FTO）是一种与肥胖风险密切相关的新基因。除了体内脂肪量和脂肪细胞大小减少外，缺乏FTO还可能导致生长迟缓。为了研究Fto基因在前脂肪细胞增殖和分化中的潜在作用，我们制造了Fto基因敲低和过表达的3T3-L1细胞。通过大量的增殖实验，我们的研究结果表明，与对照组相比，Fto敲低导致增殖抑制，线粒体膜电位降低，细胞ATP减少，细胞内脂滴减少和变小（p < 0.05）。Western blot分析表明，Fto敲低可显著抑制过氧化物酶体增殖体激活受体γ (PPARγ)和葡萄糖转运蛋白4 （GLUT4）的表达，抑制Akt磷酸化。相反，过表达Fto对细胞增殖、线粒体膜电位、ATP生成、体外分化、Akt磷酸化、PPARγ和GLUT4表达有相反的影响。此外，我们证明了磷酸肌肽3-激酶（PI3K）抑制剂Wortmannin可以抑制Fto过表达的3T3-L1细胞中的磷酸akt。综上所述，Fto通过多种机制调节3T3-L1细胞的增殖和分化，包括PPARγ和PI3K/Akt信号。
The highly regulated differentiation and proliferation of pre-adipocytes play a key role in the initiation of obesity. Fat mass and obesity associated (FTO) is a novel gene strongly associated with the risk of obesity. A deficiency of FTO may cause growth retardation in addition to fat mass and adipocyte size reduction in vivo. To investigate the potential role of Fto gene on the proliferation and differentiation of pre-adipocytes, we generated Fto-knockdown and overexpressed 3T3-L1 cells. Using numerous proliferation assays our results suggest that Fto knockdown leads to suppression of proliferation, lower mitochondrial membrane potential, less cellular ATP, and decreased and smaller intracellular lipid droplets compared with controls (p < 0.05). Western blot analysis demonstrated that Fto knockdown can significantly suppress peroxisome proliferator-activated receptor gamma (PPARγ) and glucose transporter type 4 (GLUT4) expression and inhibit Akt phosphorylation. By contrast, overexpression of Fto had the opposing effect on proliferation, mitochondrial membrane potential, ATP generation, in vitro differentiation, Akt phosphorylation, and PPARγ and GLUT4 expression. Moreover, we demonstrated that Wortmannin, a phosphoinositide 3-kinase (PI3K) inhibitor, could inhibit phospho-Akt in Fto overexpressed 3T3-L1 cells. Taken together, the results suggest that Fto regulates the proliferation and differentiation of 3T3-L1 cells via multiple mechanisms, including PPARγ and PI3K/Akt signaling.