Acta Path. Jap. 29(3): 409-411,

1979

THROMBOPLASTIC AND FIBRINOLYTIC ACTIVITIES I N VARIOUS ORGANS OF CONGENITALLY ATHYMIC NUDE MICE Seiji NAITOand Kenzo TANAKA Department of Pathology, Faculty of Nedicine, K y w h u Univerelty (Received on Sept. 26, 1978)

Thromboplastic and Abrinolytic activities in various organs of congenitally athymic nude mice were estimated. These activities varied from one organ to another and the organs were divided into six groups of possible combinations of these activities. The lung revealed high thromboplastic and high Abrinolytic activities, while the liver showed low thromboplastic and low Abrinolytic activities. The pattern of distribution of these activites in various organs of nude mice was almost similar to that of rats and mice with normal thymus. No plasmin activity was found in all organs. ACTA PATH. JAP. 29: 409-411, 1979.

Introduction. Congenitally athymic nude mice are useful animals for studying the mechanism of tumor metastasis. Recently, the role of coagulation-fibrinolysis system in tumor metastasis has attracted much attention, and thromboplastic and fibrinolytic activities of several cultured human cancer cell lines have been reported4l8. However, it is also important to know these activities in organs of nude mice, when the mechanism of metastasis is studied using these cancer cell lines and nude mice. The purpose of this report is to investigate these enzymatic properties in various organs of nude mice.

Materials and Methods Mice: Six to nine week-old male nude mice on a BALB/C background were used. Aaaay of activities: Thromboplastic and fibrinolytic activities of lung, spleen, renal medulla, renal cortex, heart, liver and brain containing meninx were estimated. Thromboplastic activity was estimated by modified Astrup's method* and was recorded with rabbit plasma recalcification time. Fibrinolytic activity was extracted by Astrup's methods and aaaayed by human plasminogen-rich fibrin agar plates prepared by the method of Nilsmn et al.@ Diameter of lysed zone was compared with that of standard urokinase (UK) (Green Cross Co., LTD., Japan) and expressed by International units of UK per one milliliter of tissue extract (IU/ml) or per one milligram of protein of tissue extract (IU/mg) determined by the method of Lowry et al'. Plasminogen-independent protease was examined with plasminogen-free fibrin agar plates in the same manner.

A@ %=,

m+ BB

Correspondence: Seiji NAITO, Department of Pathology, Faculty of Medicine, Kyushu University, 3-1-1 Maidashi, Higaehi-ku, Fukuoka 812, Japan. 409

410

THROMBOPLASTIC AND BIBRINOLYTIC ACTIVITIES IN NUDE MICE

Acta Path. Jap.

Rt?SUlts

The results are summarized in Table 1. The organs were divided into six groups of possible combinations of these activities as shown in Table 2. The lung revealed high thromboplastic and high fibrinolytic activities, while the liver showed low thromboplastic and low fibrinolytic activities. The renal medulla demonstrated moderate thromboplastic and high fibrinolytic activities, while the cortex showed moderate thromboplastic and moderate fibrinolytic activities. The brain containing meninx showed low thromboplastic and moderate fibrinolytic activities. The spleen and heart indicated moderate thromboplastic and low fibrinolytic acitivities. No plasmin activity was found in all organs. Table 1. Thromboplastic and FtWnolytic Activities in Variolle Organa of Nude Mice Organ

Thromboplastic activity (n=10) (sec*)

Lung Spleen Renal medulla Renal cortex Heart Liver Brain

l4.6fl. 1 ai.r*s. s aa. 9 f i . 6 a4.7fa.6 37.7f4.4 41.943.4 46.8f4.1

Fibrinolytic activity (n=5) (IU/ml*+)

(IU/mg***)

1.06f0.17

0.63f0.17 0.03f0.01 0. 3 7 f 0 . 3 1 0.14f0.07 0. oafo. 01 0 0 . 4 s f o . ia

~~

0.07f0.04 0.56f0.15

0.a4k0.05 0.04f0.01 0 0.36f0.08

* Activity is recorded with rabbit plasma recalcification time. ** Activity is expressed by International units of urokinase per one milliliter of

***

tissue extract. Activity is expressed by International units of urokinase per one milligram of protein of tissue extract. Table 2. Pattern of Distribution of Thrdoplastic and Fibrinolytic Activities in Organa of Nude Nice Thromboplastic activity*

-

Fibrinolytic activity** High (>0.6 IU/ml) Moderatd (0.1 0.5 IU/ml) Low (0-0.1 IU/ml)

-

High (40 seo)

Renal medulla Renal cortex Spleen, Heart

* Activity is recorded with rabbit plasma recalcification time. ** Activity is expressed by International units of urokinase per

Brain Liver one milliliter

of tissue extract.

Discussion

Thromboplastic and fibrinolytic activities in various organs of several animal species have been investigated and these activities varied from one organ to another in the same animal species and from species to species1?. The present results showed that

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these activities in various organs of nude mice varied from one organ to another. But the pattern of distribution of these activities was almost similar to that of rats and mice with normal thymus112. It was noted that thromboplastic and fibrinolytic activities of rat ascites tumor cells themsevles influenced the lodgement of tumor cells in blood-borne metastasis by aggregation of platelets and deposition of fibrin in the lungs~6~10.On the other hand, plasminogen tissue activator and tissue thromboplastin are considered to localize mainly in the endothelial cells of blood vessels and to be significant factors for formation and resolution of thrombillj12. These activities in various organs of nude mice, as shown here, may be also considered to play a significant role in the lodgement of tumor cells in the target organs. These results may be useful for studying the role of coagulation-fibrinolysis system in blood-borne metastasis using nude mice. References O.K. : The fibrinolytic activity of animal tissues. Acta Physiol. Scand. 39: 1. ALBRECHTSEN, 284-290, 1975. 2. ASTRUP, T.: Assay and content of tissue thromboplastin in different organs. Thromb. Diath. Haemorrh. 14: 401416, 1965. O.K.: Estimation of the plasminogen activator and the 3. ASTRUP,T. and ALBRECHTSEN, trypsin inhibitor in animal and human tissues. Scand. J. Clin. Lab. Invest. 9:233-243, 1957. Y., and 4. KINJO,M., OKA,K., NAITO,S., KOHGA,S., TANAKA,K., OHBOSHI,S., HAYATA,

5.

6. 7. 8.

9.

10.

11.

YASUMOTO, K. : Thromboplastic and fibrinolytic activities of cultured human cancer cell lines. Igaku-no-Ayumi 103: 148-150, 1977. (in Japanese) KINJO, M.: Lodgement and extravasation of tumor cells in blood-borne metastasis: an electron microscopic study. Br. J. Cancer 38, 1978 (in press). KOHGA,S.: Thromboplastic and fibrinolytic activities of ascites tumor cells of rats, with reference to their role in metastasis formation. Gann 69, 461470, 1978. LOWRY,O., ROSEBROUQH, N., FARR,A., and RANDALL, R.: Protein measurement with folin phenol reagent. J. Biol. Chem. 193: 265-275, 1951. NAITO,S., KINJO, M., KOHQA,S., O n , K., TANAKA, K., OHBOSHI,S., HAYATA,Y., and YASUMOTO, K. : Thromboplastic and fibrinolytic activity of cultured human cancer cells and their fate in the lung of nude mice. Cancer & Chemotherapy 5. Suppl. I: 89-95, 1978 (in Japanese). NILSSON, U.R., TOMAR,R.H., and TAYLOR,F.B.: Additional studies on human C5: development of a modified purification method and characterization of the purified product by polyacrylamide gel electrophoresis. Immunochem. 9 : 709-723, 1972. TANAKA, K., KOHQA, S., KINJO,M., and KODAMA, Y.: in Stansly, P.G. and Sato, H. (Eds.), G A " Monograph on Cancer Research: Tumor metastasis and thrombosis, with special reference to thromboplastic and fibrinolytic activities of tumor cells. No. 20, Cancer Metastasis, 97-119. University of Tokyo Press, Tokyo, 1977. TODD,A.S.: The histological localization of fibrinolysin activator. J. Path. Bact. 78: 281-

283, 1959. 12. ZELDIS, S.M., NEMERSON,Y.,

and PITLICK, F.A. : Tissue factor (Thromboplastin): Localization to plasma membranes by peroxidase-conjugated antibodies. Science 115, 70s768. 1971.

Thromboplastic and fibrinolytic activities in various organs of congenitally athymic nude mice.

Acta Path. Jap. 29(3): 409-411, 1979 THROMBOPLASTIC AND FIBRINOLYTIC ACTIVITIES I N VARIOUS ORGANS OF CONGENITALLY ATHYMIC NUDE MICE Seiji NAITOand...
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