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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2002 Oct;93(10):1090–1099. doi: 10.1111/j.1349-7006.2002.tb01210.x

Enhanced Expression of Type IV Collagen‐binding Protein (p29) in Fyn‐transfected Murine Fibrosarcoma Cells

Kazuhiko Koike 1, Katsuhisa Kogawa 1, Tetsuji Takayama 1, Naohito Yoshizaki 1, Hirohito Muramatsu 1, Kiminori Nakamura 1, Sumio Sakamaki 1, Yoshiro Niitsu 1,
PMCID: PMC5926876  PMID: 12417038

Abstract

We investigated the mechanism of the enhancement of metastatic potential induced by transfection of the fyn gene, a member of the src family. We employed two murine fyn cDNA‐transfected clones, ML‐SN1 and ML‐SN2, which were previously established from an ML‐01 low‐metastatic clone of Meth A sarcoma of BALB/c mice and were proven to have higher metastatic ability than ML‐01 and the mock‐transfected clone ML‐MT‐neo (Takayama et al., 1993). Our present investigation revealed that the two transfectants showed higher metastatic ability and higher rates of adherence to type IV collagen than ML‐MT‐neo. However, no difference was found in in vitro or in vivo growth rates, attachment to laminin or endothelial cells or cell motility through a reconstituted basement membrane. Analysis of surface membrane proteins labeled with 125I on SDS‐PAGE showed that a 29 kD band specifically bound to type IV collagen‐coupled beads was more intense in ML‐SN2 than in ML‐MT‐neo. Genistein, a protein tyrosine kinase inhibitor, dramatically reduced protein tyrosine kinase (PTK) activity of ML‐SN2 in a dose‐dependent fashion, corresponding to the reduction of adhesiveness to type IV collagen. The expression of the type IV collagen‐binding protein (p29) of ML‐SN2 was also reduced significantly by genistein treatment. These results suggested that the fyn product in Meth A cells augments the expression of a type IV collagen‐binding protein through elevation of the PTK activity of the membrane fraction and thus facilitates the metastasis of Meth A.

Keywords: fyn, Attachment, Type IV collagen, Metastasis, Meth A

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REFERENCES

  • 1. ) Fidler , I. J.Special Lecture: Critical factors in the biology of human cancer metastasis: Twenty‐eight G. H. A. Clowes Memorial Award Lecture . Cancer Res. , 50 , 6130 – 6138 ( 1990. ). [PubMed] [Google Scholar]
  • 2. ) Mogi , Y. , Kogawa , K. , Takayama , T. , Yoshizaki , N. , Bannai , K. , Muramatsu , H. , Koike , K. , Kohgo , Y. , Watanabe , N. and Niitsu , Y.Platelet aggregation induced by adenosine diphosphate released from cloned murine fibrosarcoma cells is positively correlated with the experimental metastatic potential of the cells . Jpn. J. Cancer Res. , 82 , 192 – 198 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. ) Kogawa , K. , Mogi , Y. , Takayama , T. , Koike , K. , Yoshizaki , N. , Muramatsu , H. and Niitsu , Y.Augmented expression of a type IV collagen‐binding protein in a highly metastatic murine fibrosarcoma clone . Jpn. J. Cancer Res. , 84 , 549 – 557 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Takayama , T. , Mogi , Y. , Kogawa , K. , Yoshizaki , N. , Muramatsu , H. , Koike , K. , Semba , K. , Yamamoto , T. and Niitsu , Y.A role for the fyn oncogene in metastasis of methylcholanthrene‐induced fibrosarcoma A cells . Int. J. Cancer , 54 , 875 – 879 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Heussen , C. and Dowdle , E. B.Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates . Anal. Biochem. , 102 , 196 – 202 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Fidler , I. J.Metastasis: quantitative analysis of distribution and fate of tumor emboli labeled with 125I‐5‐iodo‐2′‐deoxyuridine . J. Natl. Cancer Inst. , 45 , 773 – 782 ( 1970. ). [PubMed] [Google Scholar]
  • 7. ) Kurkinen , M. , Taylor , A. , Garrels , J. I. and Hogan , B. L. M.Cell surface‐associated proteins which bind native type IV collagen or gelatin . J. Biol. Chem. , 259 , 5915 – 5922 ( 1984. ). [PubMed] [Google Scholar]
  • 8. ) Tao , T.‐W. and Johnson , L. K.Altered adhesiveness of tumor cell surface variants with reduced metastasizing capacity‐reduced adhesiveness to vascular wall to components in culture . Int. J. Cancer , 30 , 763 – 766 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Yoshizaki , N. , Mogi , Y. , Muramatsu , H. , Koike , K. , Kogawa , K. and Niitsu , Y.Suppressive effect of recombinant human Cu, Zn‐superoxide dismutase on lung metastasis of murine tumor cells . Int. J. Cancer , 57 , 287 – 292 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Usui , H. , Yoshikawa , K. , Imazu , M. , Tsukamoto , H. and Takeda , M.Comparison of tyrosine phosphorylation of proteins by membrane fractions from mouse liver, Ehrlich ascites tumor and MH134 hepatoma . FEES Lett. , 184 , 60 – 64 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Mustelin , T. , Coggeshall , K. M. , Isakov , N. and Altman , A.T cell antigen receptor‐mediated activation of phospholipase C requires tyrosine phosphorylation . Science , 247 , 1584 – 1587 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Martin , G. R. and Timple , R.Laminin and other basement membrane components . Annu. Rev. Cell Biol. , 3 , 57 – 85 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Ura , H. , Daniel , B. R. , Reich , R. , Reddel , R. , Pfeifer , A. , Harris , C. C. and Klein‐Szanto , A. J. P.Expression of type IV collagenase and procollagen genes and its correlation with the tumorigenic, invasive, and metastatic abilities of oncogene‐transformed human bronchial epithelial cells . Cancer Res. , 49 , 4615 – 4621 ( 1989. ). [PubMed] [Google Scholar]
  • 14. ) Woessner , J. F. J.Matrix metalloproteinase and their inhibitors in connective tissue remodeling . FASEB J. , 5 , 2145 – 2154 ( 1991. ). [PubMed] [Google Scholar]
  • 15. ) Partin , A. W. , Isaacs , J. T. , Treiger , B. and Coffey , D. S.Early cell motility changes associated with an increase in metastatic ability in rat prostatic cancers cells transfected with the v‐Harvey ras oncogene . Cancer Res. , 48 , 6050 – 6053 ( 1988. ). [PubMed] [Google Scholar]
  • 16. ) Hynes , R. O.Integrins: versatility, modulation, and signaling in cell adhesion substrates . Cell , 69 , 11 – 25 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Akiyama , T. , Ishida , J. , Nakagawa , S. , Ogawara , H. , Watanabe , S. , Itoh , N. , Shibuya , M. and Fukami , Y.Genistein, a specific inhibitor of tyrosine‐specific protein kinases . J. Biol. Chem. , 262 , 5592 – 5595 ( 1987. ). [PubMed] [Google Scholar]
  • 18. ) Linassier , C. , Pierre , M. , Le Pecq , J.‐B. and Pierre , J.Mechanisms of action in NIH3T3 cells of Genistein, an inhibitor of EGF receptor tyrosine kinase activity . Biochem. Pharmacol. , 39 , 187 – 193 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Semba , K. , Nishizawa , M. , Miyazima , M. , Yoshida , M. C. , Sukegawa , J. , Yamanashi , Y. , Sasaki , M. , Yamamoto , T. and Toyoshima , K.yes‐related protooncogene, syn, belongs to the protein‐tyrosine kinase family . Proc. Natl. Acad. Sci. USA , 83 , 5459 – 5463 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. ) Kawakami , T. , Pennington , C. Y. and Robbibs , K. C.Isolation and oncogenic potential of a novel human ire‐like gene . Mol. Cell. Biol. , 6 , 4195 – 4201 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Cooke , M. P. , Abraham , K. M. , Forbush , K. A. and Perlmutter , R. M.Regulation of T‐cell receptor signaling by a src family protein‐tyrosine kinase (p59fyn) . Cell , 65 , 281 – 291 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Samelson , L. E. , Phillips , A. F. , Luong , E. T. and Klausner , R. D.Association of the fyn protein‐tyrosine kinase with the T‐cell antigen receptor . Proc. Natl. Acad. Sci. USA , 87 , 4358 – 4362 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Takeuchi , M. , Kuramochi , S. , Fusaki , N. , Nada , N. , Kawamura‐Tsuzuku , J. , Matsuda , S. , Semba , K. , Toyoshima , K. , Okada , M. and Yamamoto , T.Functional and physical interaction of protein‐tyrosine kinases fyn and csk in the T‐cell signaling system . J. Biol. Chem. , 268 , 27413 – 27419 ( 1993. ). [PubMed] [Google Scholar]
  • 24. ) Schaller , M. D. , Borgman , C. A. , Cobb , B. S. , Vines , R. R. , Reynolds , A. B. and Parsons , J. T.pp125FAK, a structurally distinctive protein‐tyrosine kinase associated with focal adhesions . Proc. Natl. Acad. Sci. USA , 89 , 5192 – 5196 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Cobb , B. S. , Schaller , M. D. , Leu , T. H. and Parsons , J. T.Stable association of pp60src and pp59fyn with the focal adhesion‐associated protein tyrosine kinase, pp125FAK . Mol. Cell. Biol. , 14 , 147 – 155 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. ) Kanazawa , S. , Ilic , D. , Hashiyama , M. , Noumura , T. , Yamamoto , T. , Suda , T. and Aizawa , S.p59fyn‐p125FAK cooperation in development of CD4+CD8+ thymocytes . Blood , 87 , 865 – 870 ( 1996. ). [PubMed] [Google Scholar]
  • 27. ) Senior , R. M. , Hinek , A. , Griffin , G. L. , Pipoly , D. J. , Crouch , E. C. and Mechem , R. P.Neutrophils show chemotaxis to type IV collagen and its 7S domain and contain a 67 kD type IV collagen binding protein with lectin properties . Am. J. Respir. Cell Mol. Biol. , 1 , 479 – 487 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Lu , M. L. , Beacham , D. A. and Jacobson , B. S.The identification and characterization of collagen receptors involved in Hela cell‐substratum adhesion . J. Biol. Chem. , 264 , 13546 – 13558 ( 1989. ). [PubMed] [Google Scholar]
  • 29. ) Hynes , R. O.Cell adhesion: old and new questions . Trends Cell Biol. , 12 , M33 – M37 ( 1999. ). [PubMed] [Google Scholar]
  • 30. ) Guan , J. L.Role of focal adhesion kinase in integrin signaling . Int. J. Biochem. Cell Biol. , 29 , 1085 – 1096 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Staunton , D. E. , Martin , S. D. , Stratowa , C. , Dustin , M. L. and Springer , T. A.Primary structure of ICAM‐1 demonstrates interaction between members of the immunoglobulin and integrin supergene families . Cell , 52 , 925 – 933 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Mattila , P. , Majuri , M.‐L. and Renkonen , R.VLA‐4 integrin on sarcoma cell lines recognizes endothelial VCAM‐1. Differential regulation of the VLA‐4 activity on various sarcoma cell lines . Int. J. Cancer , 52 , 918 – 923 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Ruoslahti , E.Fibronectin and its receptors . Annu. Rev. Biochem. , 57 , 375 – 413 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Rao , C. N. , Margulies , I. M. K. , Tralka , T. S. , Terranova , V. P. , Madri , J. A. and Liotta , L. A.Isolation of a subunit of laminin and its role in molecular structure and tumor cell attachment . J. Biol. Chem. , 257 , 9740 – 9744 ( 1982. ). [PubMed] [Google Scholar]

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