Thrombin stimulates arachidonate metabolism in murine tumor cells

M. G. Lampugnani, M. B. Donati

Research output: Contribution to journalArticle

Abstract

Thrombin can be formed in the tumor cell microenvironment following activation of the clotting cascade by procoagulants of cancer or host cells. We have tested here the effects of thrombin, either 'endogenous' or 'exogenous' (see below), on arachidonate mobilization from membrane phospholipids of mouse mammary tumor virus-induced (MMTV) carcinoma cells. These tumor cells exhibit in vitro a tissue type procoagulant activity (130 thromboplastin units/104 cells) and are therefore able to induce thrombin formation in a plasmatic milieu. To verify the effect of thrombin formation by tumor cell procoagulant ('endogenous thrombin'), either human or mouse platelet-free plasma (20% in DMEM) was added to the cell layer (prelabelled for 5 hr with a trace amount (0.013 μM) of 3H-arachidonate) and the system was recalcified (15 mM CaCl2). Thin-layer radiochromatography of the culture medium showed a significant release of 3H-labelled arachidonate products PGE2, PGF2α and 6-ketoPGFIα after 1 hr of incubation. To verify the effect of thrombin formation from host sources ('exogenous thrombin'), either bovine or purified human α-thrombin (0.1-10 U/ml) was added to the cells for different periods (from 5 min to 20 hr). Exogenous thrombin stimulated arachidonate release and metabolism in a dose-related manner. With short labelling periods (.013 μM 3H-arachidonate for 30 min-1 hr) thrombin stimulated the release of unmetabolized 3H-arachidonate, but not of 3H-arachidonate metabolites. These processes were inhibited by a specific inhibitor of thrombin enzymatic activity (α-NAPAP, 140 μM) and by a cyclo-oxygenase inhibotor (ASA 4mM). Tumor-associated procoagulants may thus contribute not only to fibrin deposition but also to generation of multipotent mediators such as arachidonate metabolites.

Original languageEnglish
Pages (from-to)367-372
Number of pages6
JournalInternational Journal of Cancer
Volume39
Issue number3
Publication statusPublished - 1987

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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