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DOI: 10.1055/s-0037-1614129
Mice Deficient in Hepsin, a Serine Protease, Exhibit Normal Embryogenesis and Unchanged Hepatocyte Regeneration Ability
The authors thank Dr. Li-Tzu Li, Ms Chin-Chin Huang, and Mr. Hsiang-Ming Wang for helpful discussions and technical assistance. Many thanks to Ms Su-Jen Hsu and the clinical chemistry laboratory at the NTUH for providing assistance to blood chemistry analyses. This work is supported by grants to Dr. S. W. Lin from the National Science Counsel (no. NSC89-2318-B002-006-M51), the National Health Research Institute (no.DOH88-HR-811 and NHRIGT-EX89) and The Istitute of Biomedical Sciences, Academia Sinica, Taiwan (no. IBMS-CRC86-T06, -CRC87-T05, and -CRC88-T09).Publikationsverlauf
Received
01. Mai 2000
Accepted after revision
31. Mai 2000
Publikationsdatum:
13. Dezember 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200005/lookinside/thumbnails/10-1055-s-0037-1614129-1.jpg)
Summary
Hepsin, a liver-enriched novel serine protease, has been implicated in participating with normal cell growth, embryogenesis, and blood coagulation pathway. To study its function in vivo, we have disrupted the mouse hepsin gene by homologous recombination. Targeted disruption of the hepsin gene and ablation of hepsin message were demonstrated by Southern blotting, Northern blotting and RT-PCR analysis. Homozygous hepsin −/− mice were viable, fertile, and exhibited no gross abnormalities, as judged by the size, weight and blood coagulation (PT) assays. However, the serum concentration of the bone form of alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase of the hepsin −/− mice was mildly elevated, in spite of no obvious pathological change of hepatocytes. To examine whether hepsin is involved in controlling cell growth in adult tissues, 70% hepatectomy was applied to the hepsin −/− mice. Liver regeneration proceeded normally in the hepsin −/− mice as judged by the liver mass restoration rate. These results suggest that loss of hepsin function causes no effect in cell growth and embryogenesis in vivo, which is in contradiction to the studies using in vitro cell culturing system. Moreover, gross mass regeneration of liver after damage proceeds normally in the absence of functional hepsin.
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References
- 1 Kurachi K, Torres-Rosado A, Tsuji A. Hepsin. Methods Enzymology 1994; 244: 100-14.
- 2 Leytus SP, Loeb KR, Hagen FS, Kurachi K, Davie EW. A Novel trypsinlike serine protease (hepsin) with a putative transmembrane domain expressed by human liver and hepatoma cells. Biochemistry 1988; 27: 1067-74.
- 3 Vu TKH, Liu RW, Haakasma CJ, Tomasek JJ, Howard EW. Identification and cloning of the membrane-associated serine protease, hepsin, from mouse preimplantation embryos. J Biol Chem 1997; 272: 31315-20.
- 4 Tsuji A, Torres-Rosado A, Arai T, Le Beau MM, Lemons RS, Chou SH, Kurachi K. Hepsin, a cell membrane-associated protease. J Biol Chem 1991; 266: 16948-53.
- 5 Tanimoto H, Yan Y, Clarke J, Korourian S, Shigemasa K, Parmley TH, Parham GP, O’Brien TJ. Hepsin, a cell surface serine protease identified in hepatoma cells, is overexpressed in ovarian cancer. Cancer Res 1997; 15: 2884-7.
- 6 Zacharski LR, Ornstein DL, Memoli AV, Rousseau SM, Kisiel W. Expression of the factor VII activating protease, Hepsin, in situ in renal cell carcinoma. Thromb Haemost 1998; 79: 876-7.
- 7 Rapaport SI, Rao LVM. The tissue factor pathway: how it has become a »Prima Ballerina«. Thromb Haemost 1995; 74: 7-17.
- 8 Torres-Rosado A, O’Shea KS, Tsuji A, Chou SH, Kurachi K. Hepsin, a putative cell-surface serine protease, is required for mammalian cell growth. Proc Natl Acad Sci 1993; 90: 7181-5.
- 9 Kawamura S, Kurachi S, Deyashiki Y, Kurachi K. Complete nucleotide sequence, origin of isoform and functional characterization of the mouse hepsin gene. Eur J Biochem 1999; 262: 755-64.
- 10 Chou PY, Fasman GD. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymed Relat Areas Mol Biol 1978; 47: 35-41.
- 11 Kazama Y, Hamamoto T, Foster DC, Kisiel W. Hepsin, a putative membrane-associated serine protease, activates human factor VII and initiates a pathway of blood coagulation on the cell surface leading to thrombin formation. J Biol Chem 1995; 270: 66-72.
- 12 Yoshida E, Verrusio EN, Mihara H, Oh D, Kwaan HC. Enhancement of the expression of urokinase-type plasminogen activator from PC-3 human prostate cancer cells by thrombin. Cancer Res 1995; 54: 3300-4.
- 13 Wojtukiewica MZ, Tang DG, Ciarelli JJ, Nelson KK, Walz DA, Diglio CA, Mammen EF, Honn KV. Thrombin increases the metastatic potential of tumor cells. Int J Cancer 1993; 54: 793-806.
- 14 Nierodzik MLR, Kajumo F, Karpatkin S. Effect of thrombin treatment of tumor cells on adhesion of tumor cells to platelets in vitro and tumor metastasis in vivo. Cancer Res 1992; 52: 3267-72.
- 15 Nierodzik MLR, Plotkin A, Kajumo F, Karpatkin S. Thrombin stimulates tumor-platelet adhesion in vitro and metastasis in vivo. J Clin Invest 1991; 87: 229-36.
- 16 Klepfish A, Greco MA, Karpatkin S. Thrombin stimulates melanoma tumor-cell binding to endothelial cells and subendothelial matrix. Int J Cancer 1993; 53: 978-8223.
- 17 Brown JM. A study of the mechanism by which anticoagulant with warfarin inhibits blood-bone metastasis. Cancer Res 1973; 33: 1217-24.
- 18 Fischer EG, Ruf W, Mueller BM. Tissue factor-initiated thrombin generation activates the signaling thrombin receptor on malignant melanoma cells. Cancer Res 1995; 55: 1629-32.
- 19 Wu Q, Yu D, Post J, Halk-Miller M, Sadler E, Morser J. Generation and characterization of mice deficient in hepsin, a hepatic transmembrane serine protease. J Clin Invest 1998; 101: 321-6.
- 20 Tybulewicz VLJ, Crawford CE, Jackson PK, Bronson RT, Mulligan RC. Neonatal lethality and lymphopenia in mice with homozygous disruption of the c-abl protooncogen. Cell 1991; 65: 1153-9.
- 21 Colucci-Guyon E, Portier MM, Dunia I, Paulin D, Pournin S, Babinet C. Mice lacking vimentin develop and reproduce without an obvious phenotype. Cell 1994; 79: 679-94.
- 22 Hogan B, Beddington R, Costantini F, Lacy E. Manipulating the mouse embryo. 2nd Edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor; New York: 1994
- 23 Joyner AL. Gene targeting-a practical guide. IRL Press; 1993
- 24 Ramirez-Solis R, Davis AC, Bradley A. Gene targeting in embryonic stem cells. Methods Enzymology 1993; 225: 878-910.
- 25 Sambrook J, Fritsch EF, Maniatis T. Molecular cloning-a laboratory manual. 2nd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor Laboratory; New York: 1989. 7: 39-42.
- 26 Chang YJ, Hamaguchi N, Chang SC, Ruf W, Shen MC, Lin SW. Engineered recombinant factor VII Q217 variants with altered inhibitor specificities. Biochemistry 1999; 38: 10940-8.
- 27 Higgins GM, Anderson RM. Experimental pathology of the liver. I. Restoration of the liver of the white rat following partial surgical removal. Arch Pathol 1931; 12: 186-202.
- 28 Wildgoose P, Nemerson Y, Hansen LL, Nielsen FE, Glazer S, Hedner U. Measurement of basal levels factor VIIa in hemophilia A and B patients. Blood 1992; 80: 25-8.
- 29 Michalopoulos GK, De Frances MC. Liver regeneration. Science 1997; 276: 60-6.
- 30 Nicolson GL. Cancer metastasis: tumor cell and host organ properties important in metastasis to specific secondary sites. Biochim Biophys Acta 1988; 948: 175-208.
- 31 Fidler IJ. Tumor heterogenity and the biology of cancer invasion and metastasis. Cancer Res 1978; 38: 2651-60.
- 32 Dvorak HF, Dickersin GR, Dvorak AM, Manseau EJ, Pyne K. Human breast carcinoma: fibrin deposits and desmoplasia inflammatory cell type and distribution, Microvasculature and infaction. J Natl Cancer Inst 1981; 67: 335-45.
- 33 Zacharski LR, Schned AR, Sorenson GD. Occurrence of fibrin and tissue factor antigen in human small cell carcinoma of the lung. Cancer Res 1983; 43: 3963-8.
- 34 Rao LVM. Tissue factor as a tumor procoagulant. Cancer metastasis Rev 1992; 11: 249-66.
- 35 Gordon SG. Cancer cell procoagulants and their role in malignant disease. Semin Thromb Hemost 1992; 18: 424-33.
- 36 Costantini V, Zacharski LR. Fibrin and cancer. Thromb Haemost 1993; 69: 406-14.
- 37 Zacharski LR, Wojtukiewicz MZ, Costantini V, Ornstein DL, Memoli VA. Pathways of coagulation/fibrinolysis activation in malignancy. Semin Thromb Haemost 1992; 18: 104-16.
- 38 Zacharski LR, Henderson WG, Rickles FR, Forman WB, Cornell CJJ, Forcier RJ. et al. Effect of warfarin anticoagulation on survival in carcinoma of the lung, colon, head and neck, and prostate; final report of VA cooperative study #75. Cancer 1984; 53: 2046-52.
- 39 Carlson SA, Ramshaw IA, Warrington RC. Involvement of plasmogen activator production with tumor metastasis in a rat model. Cancer Res 1984; 44: 3012-6.
- 40 Marutsuka K, Suzumiya J, Kataoka H, Komada N, Koono M, Sumiyoshi A. A correlation between urokinase-type plasminogen activator production and the metastatic ability of human rectal cancer cells. Invasion Metastasis 1991; 11: 181-91.