Lin F T, Lane M D
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Genes Dev. 1992 Apr;6(4):533-44. doi: 10.1101/gad.6.4.533.
Previous studies suggest that the CCAAT/enhancer-binding protein (C/EBP) functions in the coordinate expression of adipocyte genes during differentiation of 3T3-L1 preadipocytes. We sought to block expression of C/EBP selectively using a bovine papilloma virus (BPV) vector to direct transcription of a approximately 0.4-kb segment of C/EBP cDNA (in antisense orientation) containing translated sequence 5' to that encoding the basic and leucine zipper regions of the protein. Vector-directed expression of antisense C/EBP RNA in 3T3-L1 preadipocytes inhibited expression of C/EBP mRNA and protein, as well as several adipose-specific mRNAs, and also prevented cytoplasmic triglyceride accumulation. Rescue of the "adipocyte phenotype" was accomplished by transfection of cells expressing antisense RNA with a modified BPV vector that directs transcription of the complementary sense C/EBP RNA.
先前的研究表明,CCAAT/增强子结合蛋白(C/EBP)在3T3-L1前脂肪细胞分化过程中对脂肪细胞基因的协同表达起作用。我们试图利用牛乳头瘤病毒(BPV)载体选择性地阻断C/EBP的表达,以指导一段约0.4kb的C/EBP cDNA片段(反义方向)的转录,该片段包含该蛋白质基本区域和亮氨酸拉链区域编码序列5'端的翻译序列。载体介导的反义C/EBP RNA在3T3-L1前脂肪细胞中的表达抑制了C/EBP mRNA和蛋白质以及几种脂肪特异性mRNA的表达,还阻止了细胞质甘油三酯的积累。通过用一种指导互补正义C/EBP RNA转录的改良BPV载体转染表达反义RNA的细胞,实现了“脂肪细胞表型”的挽救。