Wade F Marlene, Wakade Chandramohan, Mahesh Virendra B, Brann Darrell W
Medical College of Georgia, Institute of Molecular Medicine and Genetics, 1120 15th Street, Augusta, GA 30912, USA.
Obes Res. 2005 May;13(5):818-22. doi: 10.1038/oby.2005.93.
This study used the mRNA differential display technique to identify differentially expressed genes during the process of adipogenesis in the preadipocyte cell line, 3T3-L1. 3T3-L1 cells were treated with dexamethasone, isobutyl-1-methylxanthine, and insulin to induce differentiation into mature adipocytes. Cells were collected at three time-points during differentiation: Day 0 (d0), or nondifferentiated; Day 3 (d3), during differentiation; and Day 10 (d10), >90% of the cells had differentiated into mature adipocytes. Initial studies yielded 18 potentially differentially regulated cDNA candidates (8 down-regulated and 10 up-regulated). Reverse Northern and Northern blots confirmed differential expression of six of the candidates. Four of the candidates up-regulated on d3 and d10 were identified by sequence analysis to be lipoprotein lipase, a well-known marker of adipocyte differentiation. A fifth candidate that was expressed in d0, but not d3 or d10, was identified as DRM/gremlin, a bone morphogenetic protein antagonist. Finally, a sixth candidate that was increased at d3 and d10 was identified as the peripheral benzodiazepine receptor, which has been implicated in proliferation, differentiation, and cholesterol transport in cells. This study is the first to show that peripheral benzodiazepine receptor and DRM/gremlin are expressed in preadipocyte cell lines and that they are differentially regulated during adipogenesis.
本研究采用mRNA差异显示技术,以鉴定前脂肪细胞系3T3-L1在脂肪生成过程中差异表达的基因。用 dexamethasone、异丁基-1-甲基黄嘌呤和胰岛素处理3T3-L1细胞,以诱导其分化为成熟脂肪细胞。在分化过程中的三个时间点收集细胞:第0天(d0),即未分化时;第3天(d3),处于分化过程中;以及第10天(d10),此时>90%的细胞已分化为成熟脂肪细胞。初步研究产生了18个潜在的差异调节cDNA候选基因(8个下调和10个上调)。反向Northern杂交和Northern杂交证实了其中6个候选基因的差异表达。通过序列分析鉴定出在d3和d10上调的4个候选基因是脂蛋白脂肪酶,这是一种众所周知的脂肪细胞分化标志物。一个在d0表达但在d3或d10不表达的第五个候选基因被鉴定为DRM/ gremlin,一种骨形态发生蛋白拮抗剂。最后,一个在d3和d10增加的第六个候选基因被鉴定为外周苯二氮䓬受体,其与细胞的增殖、分化和胆固醇转运有关。本研究首次表明外周苯二氮䓬受体和DRM/ gremlin在前脂肪细胞系中表达,并且它们在脂肪生成过程中受到差异调节。