Evidence for Thromboxane Biosynthesis in Established Cell Lines Derived from Human Lung Adenocarcinomas Walter C. Hubbard, Michael C. Alley, Theodore L. McLemore, et al. Cancer Res 1988;48:2674-2677. Published online May 1, 1988.
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[CANCER RESEARCH 48. 2674-2677, May 15. 1988]
Evidence for Thromboxane Biosynthesis in Established Cell Lines Derived from Human Lung Adenocarcinomas1 Walter C. Hubbard,2 Michael C. Alley, Theodore L. McLemore, and Michael R. Boyd Program Development Research Group, Developmental Therapeutics Program, Division of Cancer Treatment, National Cancer Institute, Frederick Cancer Research Facility. Frederick, Maryland 21701-1013
ABSTRACT Thromboxane B..(T\B.) is the stable nonenzymatic hydrolysis product of thromboxane A,, a substance implicated in the initiation of the facililame role of thrombocytes in the metastatic process. I \B, was isolated from protein-free culture medium of cell lines Calu-3, Calu-6, A549, and A549/ASC-1, derived from human lung adenocarcinomas. 1\B.. and other 20-carbon fatty acid cyclooxygenase products synthesized from exoge nous and endogenous arachidonic acid were identified by their character istic retention indices and fragmentation of electron-capture derivatives of unlabeled and deuterium-labeled products during combined capillary gas chromatography-mass spectrometry. I \B; comprised 2 to 6% of 20carbon fatty acid cyclooxygenase products biosynthesized from endoge nous arachidonic acid in calcium ionophore A23187-stimulated Calu-6 and A549/ASC-1 cells and 16 to 25% of these products in Calu-3 and A549 cells. The addition of 10 * M exogenous arachidonic acid to the cultured cells resulted in a 2- to 3-fold increase in TxB, and bisenoic prostanoid production with no significant alterations in the proportion of I vB; production. Prostaglandin K. and prostaglandin I-';.,,two prostanoids that can be formed either enzymatically or nonenzymatically from pros taglandin II.., accounted for >75% of isolatable 20-carbon fatty acid cyclooxygenase products synthesized from endogenous and exogenous arachidonic acid.
INTRODUCTION Fatty acid cyclooxygenase catalyzed transformation of ara chidonic acid to PGH23 and subsequent isomerization to PGE2 has been described for human lung carcinomas in vivo (1), lung carcinoma tissue in vitro (2), and established cell lines derived from human lung carcinomas in vitro (3-5). The role of PGE2 and other prostanoids in tumor metastasis, particularly in re lation to the interaction of tumor cells and host platelets, has been the subject of intensive investigation (6-14). It has been postulated that host platelets facilitate tumor metastasis by increasing tumor cell adhesiveness to other tumor cells, throm bocytes, and endothelial cells, as well as the basal lamina and, in turn, provide a milieu highly favorable for tumor cell growth and proliferation. In this report, we present evidence for the biosynthesis of TxA2, the predominant 20-carbon fatty acid cyclooxygenase product synthesized from arachidonic acid by activated thrombocytes ( 15-17), in established cell lines derived from human lung adenocarcinomas. MATERIALS
AND METHODS
Cell Culture. Established cell lines derived from lung tumor tissue explants, A549 (18, 19), and from pleural effusion, Calu-3 and Calu-6 Received 11/2/87; revised 2/2/88; accepted 2/15/88. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1Research supported, in part, by the National Cancer Institute, Department of Health and Human Services, under Contract N01-CO-23910 with Program Resources. Inc. 2 To whom requests for reprints should be addressed. 3The abbreviations used are: PGH2, prostaglandin II.; l'(.I . prostaglandin I ;, l'
