Elimination of Hydrocortisone from the Medium Enables Tissue Plasminogen Activator Gene Expression by Normal and Immortalized Nonmalignant Human Epithelial Cells
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Heli Myöhänen
Abstract
Human cervical epithelial cells transfected and immortalized with human papillomavirus type 16 DNA (HCE16/3) can be, like many other epithelial cells, normally grown in medium supplemented with epidermal growth factor, cholera toxin, hydrocortisone, insulin, transferrin, thyroid hormone and serum. We found that hydrocortisone diminished tissue plasminogen activator (tPA) production to an undetectable level. The removal of hydrocortisone increased urokinase plasminogen activator (uPA) activity within 2448 h and tPA activity within 4872 h, and converted the cells to a more elongated and fibroblastic phenotype. Upregulation of uPA mRNA was seen as early as at 3 h and of tPA mRNA within 4872 h. Higher molecular weight forms (97110 kDa) of plasminogen activators were seen in zymograms, apparently complexed with PAI-1, starting at 6 h both in the presence and absence of hydrocortisone. Immunoprecipitation with a PAI-1 monoclonal antibody confirmed that both uPA and tPA were complexed. We also studied normal diploid human bronchial epithelial cells (NHBE) and NHBE cells transformed with an adeno-12/SV40 hybrid virus (BEAS-2B). In both types of nonmalignant epithelial cells, the removal of hydrocortisone increased uPA activity. The omission of hydrocortisone increased tPA levels significantly in BEAS-2B cell cultures, and in NHBE cell cultures tPA became detectable at 72 h. No PA complexes were seen in these two cell types. We conclude that normal and immortalized nonmalignant epithelial cells produce tPA, but only if hydrocortisone is omitted in the growth medium.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- An Overview of Endosymbiotic Models for the Origins of Eukaryotes, Their ATP-Producing Organelles (Mitochondria and Hydrogenosomes), and Their Heterotrophic Lifestyle
- A Microsomal Ecdysone-Binding Cytochrome P450 from the Insect Locusta migratoria Purified by Sequential Use of Type-II and Type-I Ligands
- The Large Cytoplasmic Loop of the Glucose Transporter GLUT1 Is an Essential Structural Element for Function
- Visual Representation by Atomic Force Microscopy (AFM) of Tomato Spotted Wilt Virus Ribonucleoproteins
- Elimination of Hydrocortisone from the Medium Enables Tissue Plasminogen Activator Gene Expression by Normal and Immortalized Nonmalignant Human Epithelial Cells
- Molecular Cloning and Biochemical Characterisation of Proteases from Staphylococcus epidermidis
- Splice Variants of Human Cathepsin L mRNA Show Different Expression Rates
- Osmolytes as Modulators of Conformational Changes in Serpins
- Protective Activity of Aromatic Amines and Imines against Oxidative Nerve Cell Death
- Nitric Oxide Generation in Aqueous Solutions of Cigarette Smoke and Approaches to Its Origin
- A Recombinant scFv/Streptavidin-Binding Peptide Fusion Protein for the Quantitative Determination of the Scorpion Venom Neurotoxin AahI
