Mizushima H, Miyagi Y, Kikkawa Y, Yamanaka N, Yasumitsu H, Misugi K, Miyazaki K
Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University.
J Biochem. 1996 Dec;120(6):1196-202. doi: 10.1093/oxfordjournals.jbchem.a021541.
We previously reported a new laminin variant containing laminin gamma 2 (or B2t) chain, ladsin, which exerted prominent cell-scattering, cell-adhesion, and cell-migration activities. In the present study, this laminin was further characterized, and gene expression of its three subunits in various human tissues and cancer cell lines was examined by Northern blotting. cDNA cloning of the largest subunit of ladsin and partial amino acid sequencing of its beta (or B1) subunit revealed that ladsin was identical to laminin-5 (kalinin/epiligrin/ nicein). Among various human tissues, placenta, lung, and fetal kidney expressed high levels of mRNAs for the three subunits of laminin-5 (laminin alpha 3EPA, beta 3, and gamma 2 chains). Most gastric and squamous carcinoma cell lines constitutively expressed all of the three subunit mRNAs, while other types of carcinoma cell lines expressed one or two of them. The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and epidermal growth factor (EGF) strongly enhanced the gene expression of the three subunits, increasing 2 to 8-fold the secretion of laminin-5 from carcinoma cells into culture medium. However, TPA treatment did not increase the secretion of laminin beta 1 chain, a subunit of laminins-1, -3, and -6. The unique properties and inducibility by TPA and EGF of laminin-5 suggest that it is associated with growth and migration of cancer cells.
我们先前报道了一种新的层粘连蛋白变体,即含有层粘连蛋白γ2(或B2t)链的ladsin,它具有显著的细胞散射、细胞黏附及细胞迁移活性。在本研究中,对这种层粘连蛋白进行了进一步表征,并通过Northern印迹法检测了其三个亚基在各种人体组织和癌细胞系中的基因表达。ladsin最大亚基的cDNA克隆及其β(或B1)亚基的部分氨基酸测序表明,ladsin与层粘连蛋白-5(kalinin/epiligrin/nicein)相同。在各种人体组织中,胎盘、肺和胎儿肾脏中表达层粘连蛋白-5三个亚基(层粘连蛋白α3EPA、β3和γ2链)的mRNA水平较高。大多数胃癌和鳞状癌细胞系组成性表达所有三个亚基的mRNA,而其他类型的癌细胞系则表达其中的一个或两个。肿瘤启动子12-O-十四烷酰佛波醇-13-乙酸酯(TPA)和表皮生长因子(EGF)强烈增强了这三个亚基的基因表达,使癌细胞中层粘连蛋白-5分泌到培养基中的量增加了2至8倍。然而,TPA处理并未增加层粘连蛋白β1链(层粘连蛋白-1、-3和-6的一个亚基)的分泌。层粘连蛋白-5的独特性质以及TPA和EGF对其的诱导性表明,它与癌细胞的生长和迁移有关。