Skip to main content
PMC home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1997 May;88(5):484–491. doi: 10.1111/j.1349-7006.1997.tb00407.x

Regulation of Vimentin Expression and Protease‐mediated Vimentin Degradation during Differentiation of Human Monocytic Leukemia Cells

Koichi Honke 1,✉, Yoshinao Wada 1
PMCID: PMC5921461  PMID: 9247605

Abstract

Terminal differentiation of human monocytic leukemia THP‐1 cells is induced in vitro by 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA). We investigated the effects of TPA on the expression of vimentin during the differentiation of THP‐1 cells at both the mRNA and the protein level. On northern blotting analysis, a 2.1 kb vimentin mRNA was up‐regulated by TPA. On western blotting, small vimentin molecules with a molecular mass of approximately 40 kDa were observed in the soluble fraction and increased with TPA‐induction of cellular differentiation. Since larger, including intact, vimentin molecules were detectable at a high TPA dose, we assessed the possible existence of protease activity directed against vimentin in THP‐1 cells. With incubation of the cellular lysates of THP‐1 cells, the endogenous vimentin became increasingly smaller over time, suggesting the presence of a vimentin‐degrading protease. Phenylmethylsulfonyl fluoride inhibited this apparent protease activity against vimentin, suggesting the enzyme involved to be a serine protease. Interestingly, the protease activity was down‐regulated by TPA treatment. TPA‐treated THP‐1 cells were found to express a vimentin‐fUament network based on immunocytochemical analysis using an anti‐vimentin monoclonal antibody, V9. Taken together, these observations suggest that post‐translational mechanisms work in cooperation with transcriptional regulation to maintain the vimenti intermediate filament structure in differentiated THP‐1 cells.

Keywords: Intermediate filament, Macrophage, Phorbol ester, Protease, THP‐1

Full Text

The Full Text of this article is available as a PDF (624.2 KB).

REFERENCES

  • 1. ) Collins , S. J.HL‐60 promyelocytic leukemia cell line: proliferation, differentiation, and cellular oncogene expression . Blood , 70 , 1233 – 1244 ( 1987. ). [PubMed] [Google Scholar]
  • 2. ) Rovera , Q. , O'Brien , T. G. and Diamond , L.Induction of differentiation in human promyelocytic leukemia cells by tumor promoters . Science , 204 , 868 – 870 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Huberman , E. and Callaham , M. F.Induction of terminal differentiation in human promyelocytic leukemia cells by tumor‐promoting agents . Proc. Natl. Acad, Sci. USA , 76 , 1293 – 1297 ( 1979. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Tsuchiya , S. , Yamabe , M. , Yamaguchi , Y. , Kobayashi , Y. , Konno , T. and Tada , K.Establishment and characterization of a human acute monocytic leukemia cell line (THP‐1) . Int. J. Cancer , 26 , 171 – 176 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Tsuchiya , S. , Kobayashi , Y. , Goto , Y. , Okumura , H. , Nakae , S. , Konno , T. and Tada , K.Induction of maturation in cultured human monocytic leukemia cells by a phorbol diester . Cancer Res. , 42 , 1530 – 1536 ( 1982. ). [PubMed] [Google Scholar]
  • 6. ) Auwerx , J.The human leukemia cell line, THP‐1: a multi‐faceted model for the study of monocyte‐macrophage differentiation . Experientia , 47 , 22 – 30 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Fuchs , E. and Weber , K.Intermediate filaments: structure, dynamics, function, and disease . Annu. Rev. Bio-chem. , 63 , 345 – 382 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Klymkowsky , M. W.Intermediate filaments: new proteins, some answers, more questions . Curr. Opin. Cell Biol. , 7 , 46 – 54 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Dellagi , K. , Vainchenker , W. , Vinci , G. , Paulin , D. and Brouet , J. C.Alteration of vimentin intermediate filament expression during differentiation of human hemopoietic cells . EMBO J. , 2 , 1509 – 1514 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. ) Bernal , S. D. and Chen , L. B.Induction of cytoskeleton‐associated proteins during differentiation of human myeloid leukemic cell lines . Cancer Res. , 42 , 5106 – 5116 ( 1982. ). [PubMed] [Google Scholar]
  • 11. ) Dellagi , K. and Brouet , J. C.Alteration of vimentin intermediate filament expression during differentiation of HL‐60 and U937 human leukemic cell lines . Leuk. Res. , 8 , 611 – 616 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Giese , G. and Traub , P.Induction of vimentin synthesis in mouse myeloma cells MPC‐11 by 12‐O‐tetradecanoyl‐phorbol‐13‐acetate . Ear. J. Cell Biol. , 40 , 266 – 274 ( 1986. ). [PubMed] [Google Scholar]
  • 13. ) Ferrari , S. , Battini , R. , Kaczmarek , L. , Rittling , S. , Calabretta , B. , de Riel , J. K. , Philiponis , V. , Wei , J.‐F. and Baserga , R.Coding sequence and growth regulation of the human vimentin gene . Mol. Cell. Biol. , 6 , 3614 – 3620 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. ) Hass , R. , Giese , G. , Meyer , G. , Hartmann , A. , Dörk , T. , Köhler , L. , Resch , K. , Traub , P. and Goppelt‐Strübe , M.Differentiation and retrodifferentiation of U937 cells: reversible induction and suppression of intermediate filament protein synthesis . Eur. J. Cell Biol , 51 , 265 – 271 ( 1990. ). [PubMed] [Google Scholar]
  • 15. ) Bloemendal , H. , Willemsen , M. , Groenewoud , G. and Oomen , P.Isolation of the intermediate filament protein vimentin by chromatofocusing . FEBS Lett , 180 , 181 – 184 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Osborn , M. , Debus , E. and Weber , K.Monoclonal antibodies specific for vimentin . Eur. J. Cell Biol , 34 , 137 – 143 ( 1984. ). [PubMed] [Google Scholar]
  • 17. ) Bohn , W. , Wiegers , W. , Beuttenraüller , M. and Traub , P.Species‐specific recognition patterns of monoclonal antibodies directed against vimentin . Exp. Cell Res. , 201 , 1 – 7 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Honoré , B. , Madsen , P. , Basse , B. , Andersen , A. , Walbum , E. , Celis , J. E. and Leffers , H.Nucleotide sequence of cDNA covering the complete coding part of the human vimentin gene . Nucleic Acids Res. , 18 , 6692 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Gu , J. and Wada , Y.Aberrant expressions of decorin and biglycan genes in the carbohydrate‐deficient glycoprotein syndrome . J. Biochem. (Tokyo) , 117 , 1276 – 1279 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Chomczynski , P. and Sacchi , N.Single‐step method of RNA isolation by acid guanidinium thiocyanate‐phenol‐chloroform extraction . Anal. Biochem. , 162 , 156 – 159 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Perreau , J. , Lilienbaum , A. , Vasseur , M. and Paulin , D.Nucleotide sequence of the human vimentin gene and regulation of its transcription in tissues and cultured cells . Gene , 62 , 7 – 16 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Ramaekers , F. C. S. , Osborn , M. , Schmid , E. , Weber , K. , Bloemendal , H. and Franke , W. W.Identification of the cytoskeletal proteins in lens‐forming cells, a special epitheloid cell type . Exp. Cell Res. , 127 , 309 – 327 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Nelson , W. J. and Traub , P.Fractionation of the detergent‐resistant filamentous network of Ehrlich ascites tumor cells . Eur. J. Cell Biol , 23 , 250 – 257 ( 1981. ). [PubMed] [Google Scholar]
  • 24. ) Rittling , S. R. , Coutinho , L. , Amram , T. and Kolbe , M.AP‐l/jun binding sites mediate serum inducibility of the human vimentin promoter . Nucleic Acids Res. , 17 , 1619 – 1633 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Pieper , F. R. , van de Klundert , F. A. , Raats , J. M. , Henderik , J. B. , Schaart , G. , Ramaekers , F. C. and Bloemendal , H.Regulation of vimentin expression in cultured epithelial cells . Eur. J. Biochem. , 210 , 509 – 519 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Traub , P. and Vorgias , C. E.Involvement of the N‐termi‐nal polypeptide of vimentin in the formation of intermediate filaments . J. Cell Set , 63 , 43 – 67 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Nelson , W. J. and Traub , P.Purification and further characterization of the Ca2+‐activated proteinase specific for the intermediate filament proteins vimentin and desmin . J. Biol Chem. , 257 , 5544 – 5553 ( 1982. ). [PubMed] [Google Scholar]
  • 28. ) Shoeman , R. L. , Höner , B. , Stoller , T. J. , Kesselmeier , C. , Miedel , M. C. , Traub , P. and Graves , M. C.Human immunodeficiency virus type 1 protease cleaves the intermediate filament proteins vimentin, desmin, and glial fibrillary acidic protein . Proc, Natl, Acad. Sci. USA , 87 , 6336 – 6340 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Bories , D. , Raynal , M.‐C. , Solomon , D. H. , Darzynkiewicz , Z. and Cayre , Y.Down‐regulation of a serine protease, myeloblastin, causes growth arrest and differentiation of prorayelocytic leukemia cells . Cell , 59 , 959 – 968 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Takahashi , H. , Nukiwa , T. , Basset , P. and Crystal , R. G.Myelomonocytic cell lineage expression of the neutrophil elastase gene . J. Biol. Chem. , 263 , 2543 – 2547 ( 1988. ). [PubMed] [Google Scholar]
  • 31. ) Hanson , R. D. , Connolly , N. L. , Burnett , D. , Campbell , E. J. , Senior , R. M. and Ley , T. J.Developmental regulation of the human cathepsin G gene in myelomonocytic cells . J. Biol. Chem. , 265 , 1524 – 1530 ( 1990. ). [PubMed] [Google Scholar]
  • 32. ) Shimbara , N. , Orino , E. , Sone , S. , Ogura , T. , Takashina , M. , Shono , M. , Tamura , T. , Yasuda , H. , Tanaka , K. and Ichihara , A.Regulation of gene expression of proteasomes (multi‐protease complexes) during growth and differentiation of human hematopoietic cells . J. Biol Chem. , 267 , 18100 – 18109 ( 1992. ). [PubMed] [Google Scholar]
  • 33. ) Nakamura , N. , Tsuru , A. , Hirayoshi , K. and Nagata , K.Purification and characterization of a vimentin‐specific protease in mouse myeloid leukemia cells. Regulation during differentiation and identity with cathepsin G . Eur. J. Biochem. , 205 , 947 – 954 ( 1992. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES