Abstract
The urokinase-type plasminogen activator (uPA) interacts with its receptor (uPAR) to promote proteolysis as well as cell proliferation and migration. These functions contribute to the pathogenesis of neoplastic growth and invasiveness. Expression of uPAR in tumor extracts also inversely correlates with prognosis in many forms of cancer. In this study, we sought to determine if differences in uPAR expression were distinguishable between cultured human lung carcinoma and malignant mesothelioma subtypes. We also sought to determine if, as in malignant mesothelioma cells, uPAR expression is regulated at the posttranscriptional level in cultured malignant lung carcinoma cells. Using 125I-uPA binding and ligand blotting techniques, uPAR was expressed by phenotypically diverse lung carcinoma cell lines, including the H460, H157 and H1395 non-small cell lines and the H146 small cell lung carcinoma line. Increased uPAR expression was also detected in spindle-shaped (M33K) and epithelioid (M9K and MS-1) malignant mesothelioma cells. Selected mediators, including TGF-β, TNF-α, LPS and PMA, uniformly enhanced uPAR expression in each of the tumor cell lines. Steady state uPAR mRNA expression was determined by RNase protection assay and correlated directly with the changes in cell surface uPAR expression. By gel mobility shift and UV-cross linking assays, a uPAR mRNA binding protein (uPAR mRNABp) implicated in the posttranscriptional control of message stability, was identified in each of the cell lines. Expression of uPAR and its message in cultured lung carcinoma and malignant mesothelioma cells is similarly influenced by effectors present in the tumor microenvironment. Regulation of the uPAR message occurs at the posttranscriptional level in cultured small and non-small cell lung carcinoma cells as well as spindle-shaped and fibrous malignant mesothelioma cell lines. Posttranscriptional regulation of uPAR in all these cells involves the interaction of the uPAR mRNABp with uPAR mRNA, which promotes uPAR mRNA destabilization.
Similar content being viewed by others
References
Dano K, Andreasen PA, Grondahl-Hansen J, Kristensen P, Nielsen LS, Skriver L: Plasminogen activators, tissue degradation, and cancer. Adv Cancer Res 44: 139–266, 1985
Mignatti P, Rifkin DB: Biology and biochemistry of proteinases in tumor invasion. Physiol Rev 73: 161–195, 1993
Liotta LA, Stetler-Stevenson WG, Steeg PS: Cancer invasion and metastasis: Positive and negative regulatory elements. Cancer Invest 9: 543–551, 1991
Pedersen H, Brunner N, Francis D, Osterlind K, Ronne E, Hansen HH, Dano K, Grondahl-Hansen J: Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res 54: 4671–4675, 1994
Lund LR, Ellis V, Ronne E, Pyke C, Dano K: Transcriptional and posttranscriptional regulation of the receptor for urokinase-type plasminogen activator by cytokines and tumour promoters in the human lung carcinoma cell line A549. Biochem J 310: 345–352, 1995
Lund LR, Ronne E, Roldan AL, Behrendt N, Romer J, Blasi F, Dano K: Urokinase receptor mRNA level and gene transcription are strongly and rapidly increased by phorbol myristate acetate in human monocytelike U937 cells. J Biol Chem 266: 5177–5181, 1991
Shetty S, Kumar A, Idell S: Posttranscriptional regulation of urokinase receptor mRNA: Identification of a novel urokinase receptor mRNA binding protein in human mesothelioma cells. Mol Cell Biol 17: 1075–1083, 1997
Pepper MS, Matsumoto K, Nakamura T, Orci L, Montesano R: Hepatocyte growth factor increases urokinase-type plasminogen activator (u-PA) and u-PA receptor expression in Madin-Darby canine kidney epithelial cells. J Biol Chem 267: 20493–20496, 1992
Ganesh S, Sier CF, Heerding MM, Griffioen G, Lamers CB, Verspaget HW: Urokinase receptor and colorectal cancer survival. Lancet 344: 401–402, 1994
Duggan C, Maguire T, McDermott E, O'Higgins N, Fennelly JJ, Duffy MJ: Urokinase plasminogen activator and urokinase plasminogen activator receptor in breast cancer. Int J Cancer 61: 597–600, 1995
Grondahl-Hansen J, Hilsenbeck SG, Christensen IJ, Clark GM, Osborne CK, Brunner N: Prognostic significance of PAI-1 and uPA in cytosolic extracts obtained from node-positive breast cancer patients. Breast Cancer Res Treat 43: 153–163, 1997
Heiss MM, Allgayer H, Gruetzner KU, Funke I, Babic R, Jauch KW, Schildberg FW: Individual development and uPA-receptor expression of disseminated tumour cells in bone marrow: A reference to early systemic disease in solid cancer. Nature Med 1: 1035–1039, 1995
Shetty S, Idell S: A urokinase mRNA binding protein-mRNA interaction regulates receptor expression and function in human pleural eesothelioma cells. Arch Biochem Biophys 356: 265–279, 1998
Shetty S, Idell S: A urokinase receptor mRNA binding protein from rabbit lung fibroblasts and methothelial cells. Am J Physiol 274: 871–882, 1998
Shetty S, Kumar A, Johnson A, Pueblitz S, Idell S: Urokinase receptor in human malignant mesothelioma cells: Role in tumor cell mitogenesis and proteolysis. Am J Physiol 268: L972–L982, 1995
Shetty S, Kumar A, Johnson AR, Pueblitz S, Holiday D, Raghu G, Idell S: Differential expression of the urokinase receptor in fibroblasts from normal and fibrotic human lungs. Am J Respir Cell Mol Biol 15: 78–87, 1996
Yebra M, Parry GN, Stromblad S, Mackman N, Rosenberg S, Mueller BM, Cheresh DA: Requirement of receptor-bound urokinase-type plasminogen activator for integrin alphavbeta5-directed cell migration. J Biol Chem 271: 29393–29399, 1996
Shetty S, Kumar A, Johnson AR, Idell S: Regulation of mesothelial cell mitogenesis by antisense oligonucleotides for the urokinase receptor. Antisense Res Dev 5: 307–314, 1995
Blasi F, Vassalli JD, Dano K: Urokinase-type plasminogen activator: Proenzyme, receptor, and inhibitors. J Cell Biol 104: 801–804, 1987
Vassalli JD, Sappino AP, Belin D: The plasminogen activator/plasmin system. J Clin Invest 88: 1067–1072, 1991
Bruckner A, Filderman AE, Kirchheimer JC, Binder BR, Remold HG: Endogenous receptor-bound urokinase mediates tissue invasion of the human lung carcinoma cell lines A549 and Calu-1. Cancer Res 52: 3043–3047, 1992
Boyd D, Florent G, Kim P, Brattain M: Determination of the levels of urokinase and its receptor in human colon carcinoma cell lines. Cancer Res 48: 3112–3116, 1988
Saksela O, Rifkin DB: Cell-associated plasminogen activation: Regulation and physiological functions. Ann Rev Cell Biol 4: 93–126, 1988
Ossowski L: In vivo invasion of modified chorioallantoic membrane by tumor cells: The role of cell surface-bound urokinase. J Cell Biol 107: 2437–2445, 1988
Achbarou A, Kaiser S, Tremblay G, Ste-Marie LG, Brodt P, Goltzman D, Rabbani SA: Urokinase overproduction results in increased skeletal metastasis by prostate cancer cells in vivo. Cancer Res 54: 2372–2377, 1994
Schmitt M, Janicke F, Graeff H: Tumour-associated fibrinolysis: The prognostic relevance of plasminogen activators uPA and tPA in human breast cancer. Blood Coag Fibrinol 1: 695–702, 1990
Hollas W, Hoosein N, Chung LW, Mazar A, Henkin J, Kariko K, Barnathan ES, Boyd D: Expression of urokinase and its receptor in invasive and non-invasive prostate cancer cell lines. Thromb Haemost 68: 662–666, 1992
Mitsubayashi S, Akiyama T, Kurita T, Okada K, Bando H, Sakai T, Matsuo O: Plasminogen activator in bladder tumors. Urol Res 15: 335–339, 1987
Janicke F, Schmitt M, Graeff H: Clinical relevance of the urokinasetype and tissue-type plasminogen activators and of their type 1 inhibitor in breast cancer. Semin Thromb Hemost 17: 303–312, 1991
Hofmann R, Lehmer A, Buresch M, Hartung R, Ulm K: Clinical relevance of urokinase plasminogen activator, its receptor, and its inhibitor in patients with renal cell carcinoma. Cancer 78: 487–492, 1996
Brunner N, Pyke C, Hansen CH, Romer J, Grondahl-Hansen J, Dano K: Urokinase plasminogen activator (uPA) and its type 1 inhibitor (PAI-1): Regulators of proteolysis during cancer invasion and prognostic parameters in breast cancer. (Review) (76 refs). Cancer Treat Res 71: 299–309, 1994
Shaw G, Kamen R: A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 46: 659–667, 1986
Shyu AB, Greenberg ME, Belasco JG: The c-fos transcript is targeted for rapid decay by two distinct mRNA degradation pathways. Genes Dev 3: 60–72, 1989
Crowley CW, Cohen RL, Lucas BK, Liu G, Shuman MA, Levinson AD: Prevention of metastasis by inhibition of the urokinase receptor. Proc Nat Acad Sci USA 90: 5021–5025, 1993
Raymond V, Atwater JA, Verma IM: Removal of an mRNA destabilizing element correlates with the increased oncogenicity of protooncogene fos. Oncogene Res 5: 1–12, 1989
Ross HJ, Sato N, Ueyama Y, Koeffler HP: Cytokine messenger RNA stability is enhanced in tumor cells. Blood 77: 1787–1795
Gyetko MR, Todd RF, Wilkinson CC, Sitrin RG: The urokinase receptor is required for human monocyte chemotaxis in vitro. J Clin Invest 93: 1380–1387, 1994
Ossowski L, Clunie G, Masucci MT, Blasi F: In vivo paracrine interaction between urokinase and its receptor: Effect on tumor cell invasion. J Cell Biol 115: 1107–1112, 1991
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shetty, S., Idell, S. Posttranscriptional regulation of urokinase receptor gene expression in human lung carcinoma and mesothelioma cells in vitro. Mol Cell Biochem 199, 189–200 (1999). https://doi.org/10.1023/A:1006914800447
Issue Date:
DOI: https://doi.org/10.1023/A:1006914800447