Sakariassen K S, Powell J S, Raines E W, Ross R
Department of Pathology, University of Washington, Seattle, 98195.
J Cell Biochem. 1989 Jan;39(1):87-94. doi: 10.1002/jcb.240390110.
The genes for platelet-derived growth factor (PDGF) A and PDGF B chains are expressed in a variety of biological situations. Active PDGF consists of two distinct but homologous polypeptide chains, PDGF A and PDGF B, which are found as heterodimers or homodimers. We report a novel situation in which there is selective expression of mRNA encoding PDGF B in cell lines derived from baby hamster kidney (BHK) following transfection with various gene/cDNA constructs and following growth selection with methotrexate. The process of transfection itself, and not expression of the proteins encoded by the transfected genes/cDNAs (hormones, enzymes, and structural proteins), induces expression of PDGF B. No PDGF B mRNA is detectable in control cell lines. Low levels of mRNA encoding PDGF A are constitutively present and are not changed by transfection and or growth selection. PDGF-like activity is present in the medium whenever PDGF B mRNA is detected. The composition of the secreted PDGF dimer cannot be established from our data, but quantitative analysis of mRNA suggests that the PDGF is a B-B dimer. However, the data show that transcription of the PDGF A and PDGF B genes in BHK cells is regulated independently, similar to that reported for some human tumor cells. Furthermore, the selective expression of PDGF B in response to the introduction of foreign genes and to growth selection may be an important aspect of the reaction of these cells to nonoptimal growth conditions, allowing survival and growth of the cells that express PDGF B.
血小板衍生生长因子(PDGF)A链和B链的基因在多种生物学情况下表达。活性PDGF由两条不同但同源的多肽链组成,即PDGF A和PDGF B,它们以异二聚体或同二聚体形式存在。我们报告了一种新情况,在用各种基因/cDNA构建体转染并经甲氨蝶呤进行生长筛选后,源自幼仓鼠肾(BHK)的细胞系中存在编码PDGF B的mRNA的选择性表达。转染过程本身,而非转染基因/cDNA(激素、酶和结构蛋白)所编码蛋白质的表达,诱导了PDGF B的表达。在对照细胞系中未检测到PDGF B mRNA。编码PDGF A的mRNA以组成型方式低水平存在,并且不受转染和/或生长筛选的影响。只要检测到PDGF B mRNA,培养基中就存在PDGF样活性。从我们的数据无法确定分泌的PDGF二聚体的组成,但对mRNA的定量分析表明该PDGF是B - B二聚体。然而,数据显示BHK细胞中PDGF A和PDGF B基因的转录是独立调节的,这与一些人类肿瘤细胞的情况类似。此外,PDGF B对外源基因导入和生长筛选的选择性表达可能是这些细胞对非最佳生长条件反应的一个重要方面,使得表达PDGF B的细胞能够存活和生长。