Abstract
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.
Highlights
Obesity increasingly is a pressing public health concern with a rapidly surging prevalence in both developed and developing countries [1,2,3]
Our results demonstrate that fat mass and obesity associated gene (Fto) siRNA transfection effectively suppressed Fto at both mRNA and protein level (Figure 1A,B), while the recombinant plasmid transfection increased Fto expression in
Construction of Fto knockdown or overexpressed pre-adipocyte model: (A) 3T3-L1 cells were transfected with siRNA control or three Fto specific siRNAs for 24 h
Summary
Obesity increasingly is a pressing public health concern with a rapidly surging prevalence in both developed and developing countries [1,2,3]. According to the World Health Organization, in 2014 more than 1.9 billion adults were overweight, 600 million of whom were obese [4]. It has been widely reported that the majority of obese individuals have multiple comorbidities including inflammation, hypertension, coronary heart disease and stroke, type 2 diabetes mellitus, insulin resistance, gall bladder disease, nonalcoholic fatty liver disease, and cancers, among others. Together, these result in the poor health of those affected and may result in accelerated aging and death [3,5]. The existing interventions for obesity control, including diet modification and exercise-based managements, seem largely ineffective [5], in part related to our inadequate and Nutrients 2016, 8, 102; doi:10.3390/nu8020102 www.mdpi.com/journal/nutrients
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