Synthesis of Vitamin K-Dependent Proteins by Cultured Human Tumor Cells^[*]

Summary

The observation that warfarin inhibits the growth and metastasis of certain types of clinical and experimental tumors suggests a role for vitamin K in tumor biology. We have investigated synthesis of vitamin K-dependent proteins in four malignant (lung epidermoid carcinoma, melanoma, colon adenocarcinoma, and breast adenocarcinoma) and three normal (colon epithelium, breast epithelium, and fibroblasts) cell lines of human origin grown in tissue cultures. Our results show the following: 1) Vitamin K-dependent carboxylase activity is present in all of the malignant and normal cell lines studied. 2) The malignant as well as normal cell lines synthesize a family of vitamin K-dependent proteins. Microsomal precursors of these proteins with apparent molecular mass of 74, 62, and 34 kDa are common to all malignant and normal cell lines whereas precursors of higher and lower molecular mass seem to be synthesized by some but not all tumor cell lines. 3) The 74 kDa precursor synthesized by colon carcinoma and breast carcinoma was positively identified as a precursor of protein S.

Keywords

Vitamin K - Human cancer - Clotting factors - Tumorprocoagulant

• References

• 1 Zacharski LR. Basis for selection of anticoagulant drugs for therapeutic trials in human malignancy. Hemostasis 1986; 16: 300-320
• 2 Zacharski LR, Henderson WG, Rickles FR, Forman WG, Cornell Jr CJ, Forcier RJ, Headley E, Kim SH, O'Donnell JF, O'Dell R, Tomyos K, Kwaan HC. Effect of sodium warfarin on survival in small cell carcinoma of the lung. JAMA 1981; 145: 831-835
• 3 Hoover Jr HC, Ketcham AS, Millar RC, Gralnick HR. Osteosarcoma: improved survival with anticoagulation and amputation. Cancer 1978; 41: 2475-2480
• 4 Thornes RD. Adjuvant therapy of cancer via the cellular immune mechanism or fibrin by induced fibrinolysis and oral anticoagulants. Cancer 1975; 35: 91-97
• 5 Zacharski LR, Henderson WG, Rickies FR, Forman WB, Cornell Jr CJ, Forcier RJ, Edwards RL, Headley E, Kim SH, O'Donnell JF, O'Dell R, Tornyos K, Kwaan HC. Effect of warfarin anticoagulation on survival in carcinoma of the lung, colon, head and neck, and prostate: final report of VA Cooperative Study No. 75. Cancer 1984; 53: 2046-2052
• 6 Brown JM. A study of the mechanism by which anticoagulation with warfarin inhibits blood-borne metastases. Cancer Res 1973; 33: 1217-1224
• 7 Roncaglioni MC, Dalesandro AP B, Casali B, Vermeer C, Donati MB. β-Glutamyl carboxylase activity in experimental tumor tissues: a biochemical basis for vitamin K dependence of cancer procoagulant. Haemostasis 1986; 16: 295-299
• 8 Hauschka PV, Harron Y, Buchthal SD, Bell RG. Vitamin K-dependent processes in tumor cells. Haemostasis 1986; 16: 273-287
• 9 Hilgard P. Experimental vitamin K deficiency and spontaneous metastases. Br J Cancer 1977; 35: 891-892
• 10 Hilgard P, Schulte H, Wetzig G, Schmitt G, Schmidt CG. Oral anticoagulation in the treatment of a spontaneously metastasizing murine tumor. Br J Cancer 1977; 35: 78-85
• 11 Poggi A, Mussoni L, Kornblihtt L, Ballabio E, DeGataeno G, Donati MB. Warfarin enantiomers, anticoagulation, and experimental tumor metastasis. Lancet 1978; 1: 163-164
• 12 Esmon CT, Sadowski JA, Suttie JW. A new carboxylation reaction. J Biol Chem 1975; 250: 4744-4748
• 13 Stenflo J, Suttie JW. Vitamin K-dependent formation of gamma-carboxyglutamic acid. Annu Rev Biochem 1977; 46: 157-172
• 14 Donati MB, Semeraro N. Cancer cell procoagulants and their pharmacological modulation. Haemostasis 1984; 14: 422-429
• 15 Hilgard P. The use of oral anticoagulants in tumor therapy. In: Donati MB, Davidson JF, Garattini S. (eds) Malignancy and the Hemostatic System. Raven Press; New York: 1981: 103-111
• 16 Pan LC, Price PA. The propeptide of rat bone γ-carboxyglutamic acid protein shares homology with other vitamin K-dependent protein precursors. Proc Natl Acad Sci USA 1985; 82: 6109-6113
• 17 Charles TL, Suttie JW. Vitamin K-dependent carboxylase: subcellular location of the carboxylase and enzymes involved in vitamin K. Biochemistry 1980; 19: 1161-1167
• 18 Esmon CT, Suttie JW. Vitamin K-dependent carboxylase. Solubilization and properties. J Biol Chem 1976; 251: 6238-6243
• 19 Laemmli UK. Cleavage of structured proteins during the assembly of the head of bacteriophage T₄ . Nature 1970; 277: 680-685
• 20 Fairbanks G, Steck LS, Wallach DF H. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 1971; 10: 2606-2617
• 21 Rannels SR, Gallagher KJ, Wallin R, Rannels DE. Vitamin K-dependent carboxylation of pulmonary surfactant associated proteins. Proc Natl Acad Sci USA 1987; 84: 5952-5956
• 22 Hauschka PA. Quantitative determination of γ-carboxyglutamic acid in proteins. Anal Biochem 1977; 80: 212-223
• 23 Kuwada M, Katayama U. A high performance liquid chromatographic method for the simultaneous determination of γ-carboxyglutamic acid and glutamic acid in proteins, bone and urine. Anal Biochem 1981; 117: 259-265
• 24 Reiderer BM, Zagon IS, Goodman SR. Brain spectrin (240–235) and brain spectrin (240/235E): differential expression during mouse brain development. J Neurosci 1987; 7: 864-874
• 25 Sadowski JA, Esmon CT, Suttie JW. Vitamin K-dependent carboxylase: Requirement of the rat liver microsomal enzyme system. J Biol Chem 1970; 251: 2770-2779
• 26 Schaffner W, Weissmann C. A rapid sensitive and specific method for the determination of protein in dilute solution. Annal Biochem 1973; 56: 502-514
• 27 Fair DS, Marlar R, Levin EG. Human endothelial cells synthesize protein S. Blood 1986; 67: 1168-1171
• 28 Wallin R, Rannels SR. Identification of vitamin K-dependent carboxylase activity in lung type II cells but not in lung macrophages. Biochem J 1988; 250: 557-563
• 29 Shah DV, Engelke JA, Suttie JW. Abnormal prothrombin in the plasma of rats carrying hepatic tumors. Blood 1987; 69: 850-854
• 30 Suttie JW, Vitamin K. In: The Fat-Soluble Vitamins, Handbook of Lipid Research. (; 2). Deluca HF. (ed) Plenum Press; New York: 1978: 211-277
• 31 De Boer-Van Den Berg MAG, Vitendaal MP, Vermeer C. Direct measurement of vitamin K-dependent enzymes in various isolated and cultured tumor and non-tumor cells. Mol Cell Biochem 1987; 75: 71-76
• 32 Wallin R, Suttie JW. Vitamin K-dependent carboxylation, possible artifact of analysis due to pyridine nucleotide-dependent carboxylation. Biochem Biophys Res Commun 1980; 94: 1374-1380