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目的观察人肥胖相关全长新基因STEAP4在人前体脂肪细胞诱导分化过程中不同时段表达水平的变化,探讨STEAP4基因与脂肪细胞分化、脂质积聚的关系。方法体外培养人前体脂肪细胞,待细胞生长融合后以1-甲基-3-异丁基黄嘌呤、地塞米松、胰岛素及罗格列酮联合诱导方案,在体外诱导其分化为成熟脂肪细胞。在前体脂肪细胞向成熟脂肪细胞分化过程中观察细胞形态及脂质积聚变化;采用实时荧光定量RT-PCR技术检测诱导分化不同时段(前体,第0、4、6、8、11、14、17天)脂肪细胞中STEAP4基因mRNA的表达水平。结果STEAP4基因高表达于人前体脂肪细胞;加入脂肪细胞分化诱导剂后(第4天)表达略有上调,其后随脂肪细胞分化成熟该基因表达量呈逐渐下降趋势;至脂肪细胞完全分化成熟(第14-17天)表达量最低。STEAP4基因表达水平除在诱导分化前至第4天、第14-17天差异无统计学意义外,余各时间点的表达水平差异均有统计学意义(Pa<0.05)。结论STEAP4基因随脂肪细胞分化成熟表达逐渐下调,可促进脂肪细胞的分化成熟和脂质积聚,与肥胖的发生有关。
OBJECTIVE: To observe the expression of STEAP4, a novel full-length obesity related gene, in different periods of human adipocyte differentiation and to investigate the relationship between STEAP4 gene and adipocyte differentiation and lipid accumulation. Methods Human preadipocytes were cultured in vitro. After the cells were fused, the combination of 1-methyl-3-isobutylxanthine, dexamethasone, insulin and rosiglitazone was induced to differentiate into mature adipocytes in vitro cell. The changes of cell morphology and lipid accumulation were observed during the differentiation of precursor adipocytes into mature adipocytes. Real-time fluorescent quantitative RT-PCR was used to detect the differentiation and differentiation at different time points (precursors, 0,4,6,8,11,14 , 17 days) STEAP4 gene mRNA expression levels in adipocytes. Results The expression of STEAP4 gene was highly expressed in human preadipocytes. The expression of STEAP4 gene was slightly up-regulated after adding adipocyte differentiation inducer (day 4), then decreased gradually with the differentiation and maturation of adipocytes; Mature (days 14-17) showed the lowest level of expression. The expression level of STEAP4 was not significantly different from that of day 4 to 14-17 before differentiation (P <0.05). Conclusion The STEAP4 gene is gradually down-regulated with the differentiation and maturation of adipocytes, which can promote the maturation and accumulation of adipocytes, which is related to the occurrence of obesity.