Li Wei, Fan Jianhua, Chen Mei, Guan Shengxi, Sawcer David, Bokoch Gary M, Woodley David T
The Division of Dermatology and the University of Southern California/Norris Cancer Center, University of Southern California Keck School of Medicine, Los Angeles, California 90033, USA.
Mol Biol Cell. 2004 Jan;15(1):294-309. doi: 10.1091/mbc.e03-05-0352. Epub 2003 Oct 31.
Migration of human dermal fibroblasts (HDFs) is critical for skin wound healing. The mechanism remains unclear. We report here that platelet-derived growth factor-BB (PDGF-BB) is the major promotility factor in human serum for HDF motility on type I collagen. PDGF-BB recapitulates the full promotility activity of human serum and anti-PDGF neutralizing antibodies completely block it. Although collagen matrix initiates HDF migration without growth factors, PDGF-BB-stimulated migration depends upon attachment of the cells to a collagen matrix. The PDGF-BB's role is to provide directionality and further enhancement for the collagen-initiated HDF motility. To study the collagen and PDGF-BB "dual signaling" in primary HDF, we establish "gene cassettes" plus lentiviral gene delivery approach, in which groups of genes are studied individually or in combination for their roles in HDF migration. Focal adhesion kinase, p21(Rac,CDC42)-activated kinase and Akt are grouped into an upstream kinase gene cassette, and the four major mitogen-activated protein kinases (extracellular signal-regulated kinase 1/2, p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase 5) are grouped into a downstream kinase gene cassette. The experiments demonstrate 1) the genes' individual roles and specificities, 2) their combined effects and sufficiency, and 3) the mechanisms of their intermolecular connections in HDF migration driven by collagen and PDGF-BB.
人真皮成纤维细胞(HDFs)的迁移对皮肤伤口愈合至关重要。其机制尚不清楚。我们在此报告,血小板衍生生长因子-BB(PDGF-BB)是人类血清中促进HDF在I型胶原上运动的主要因子。PDGF-BB概括了人类血清的全部促进运动活性,而抗PDGF中和抗体则完全阻断该活性。尽管胶原基质在没有生长因子的情况下可启动HDF迁移,但PDGF-BB刺激的迁移取决于细胞与胶原基质的附着。PDGF-BB的作用是为胶原启动的HDF运动提供方向性并进一步增强其运动。为了研究原代HDF中的胶原和PDGF-BB“双重信号”,我们建立了“基因盒”加慢病毒基因递送方法,其中对基因组合分别或联合研究它们在HDF迁移中的作用。粘着斑激酶、p21(Rac,CDC42)激活的激酶和Akt被归入上游激酶基因盒,而四种主要的丝裂原活化蛋白激酶(细胞外信号调节激酶1/2、p38、c-Jun NH2末端激酶和细胞外信号调节激酶5)被归入下游激酶基因盒。实验证明了1)基因的个体作用和特异性,2)它们的联合作用和充分性,以及3)在由胶原和PDGF-BB驱动的HDF迁移中它们分子间连接的机制。