| Obesity, one of the common metabolic diseases, can be the important risk factor for many severe diseases, such as type 2 diabetes, hypertension, cardiovascular diseases and cancer. The prevalence of obesity has been significantly increasing and has grown to be a global epidemic disease in recent decade. Currently, obesity rates of children and adolescents are being increased year by year. Childhood obesity poses not only a health hazard but also a significant burden of the family and society. Totally, pediatric obesity has become a very important problem and must be resolved immediately.Obesity is a multifactorial disease. It has been widely accepted that genetic factors as a key role in the development of obesity. Currently, over 600 genes, markers and chromosomal regions have been identified as associated with or linked to human obesity phenotypes. However, these known genes cannot fully clarify the etiology and pathogenesis of obesity. Thus, identification and characterization of novel genes and proteins associated with obesity remain an important issue.C10orf116, a novel obesity-related gene, was identified by suppression subtractive hybridization (SSH) in our laboratory. Previous studies showed that C10orf116 was specific expressed in nucleus and abundantly expressed in human adipose tissue, and the C10orf116 expression was significantly upregulated in obese patients. C10orf116 gene expression was dramatically up-regulated during early differentiation of human preadipocyte differentiation. In order to further clarify the C10orf116 gene function in fat cells, we examined the effect of C10orf116 on cell proliferation, apoptosis, and insulin sensitivity. Furthermore, we screened and identified the microRNA differential expression profile in mature adipocytes of C10orf116 overexpression.In the 1st part, we examined the effect of C10orf116 on cell proliferation, and apoptosis in vitro by establishing a stable preadipocyte cell line overexpressing C10orf116. We found that: (1) the result of the MTT assay indicated that C10orf116 causes the promotion of cell population growth of 3T3-L1 preadipocytes. (2) C10orf116 can prevent apoptosis induced by serum deprivation in preadipocytes with the analysis of annexin V-FITC, both caspase-3 and caspase-8 activity, Hoechst 33342 staining.In the 2nd part, 2-Deoxy-glucose labeled by isotope 3H was used to assay glucose uptake of mature adipocytes. 3T3-L1 preadipocytes were cultured in vitro and differentiated into the adipocytes by MIX, dexamethasone, insulin. The levels of GLUT4 mRNA expression at the 0th, 4th, 8th day during differentiation were evaluated by real-time quantitative RT-PCR. The data showed that ectopic expression of C10orf116 dramatically increases insulin-stimulated glucose uptake in adipocytes, but no significant difference in basal glucose uptake. Simultaneously, real-time quantitative RT-PCR analysis for the expression of GLUT4 indicated that the expression of glucose transporter was upregulated.In the 3rd part, we screened and identified the microRNA differential expression profile in mature adipocytes of C10orf116 overexpression. Filtered microRNAs were evaluated by real-time quantitative RT-PCR. By significance analysis of microarrays based on microarray screening, 77 significant difference microRNAs were obtained. In C10orf116 gene over-expression mature fat cells, 34 up-regulated and 43 down-regulated microRNAs were observed. The most significant up-regulated microRNAs were miR-712* and miR-1944, while miR-1907, miR-374, miR-15b were significantly down-regulated. Simultaneously, real-time quantitative RT-PCR analysis for the expression of miR-1944 indicated that the expression of miR-1944 was also upregulated.In conclusion, we characterized several original features of the novel gene C10orf116 although the precise functional properties of this gene remain to be clarified and await further investigation. |
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