Synthesis of cathepsin B by cells derived from the HL60 promyelocytic leukaemia cell line

D. Burnett, J. Crocker, Andrew T M Vaughan

Research output: Contribution to journalArticle

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Abstract

Cells of the human promyelocytic HL60 line were induced to differentiate into granulocyte-like cells with dimethylsulphoxide (DMSO) or macrophage-like cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). The synthesis of Cathepsin B by these cells was studied by immunoperoxidase staining and assay of cell lysates using the fluorimetric substrate benzoyloxycarbonyl-phenylanalyl-arginine-4-methyl-7-coumarylamide. Only 2-5% of the uninduced HL60 cells and DMSO-induced cells were immunohistochemically positive for Cathepsin B, compared with over 80% of the TPA-induced cells. Cathepsin B activity was lowest in the lysates of uninduced HL60s. DMSO-induced cells contained 1.5-2-fold the enzyme activity of HL60s and TPA-induced cell lysates demonstrated 5-14-fold the activity of uninduced HL60s. Induction of Cathepsin B synthesis was therefore associated with differentiation of the promyelocytes into cells of the monocyte/macrophage type, but not granulocyte-like cells. Cathepsin B was located immunohistochemically in human palatine tonsils. The enzyme was only demonstrated within macrophages in these tissues. Cathepsin B may therefore be a useful immunohistochemical marker for malignant and nonmalignant cells of the monocyte/macrophage lineage.

Original languageEnglish (US)
Pages (from-to)249-254
Number of pages6
JournalJournal of Cellular Physiology
Volume115
Issue number3
StatePublished - 1983
Externally publishedYes

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Cathepsin B
Leukemia
B-Lymphocytes
Macrophages
Cells
Cell Line
Dimethyl Sulfoxide
Acetates
Tetradecanoylphorbol Acetate
Enzyme activity
Granulocytes
Monocytes
Assays
Tissue
Granulocyte Precursor Cells
HL-60 Cells
Palatine Tonsil
Enzymes
Substrates
Staining and Labeling

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Synthesis of cathepsin B by cells derived from the HL60 promyelocytic leukaemia cell line. / Burnett, D.; Crocker, J.; Vaughan, Andrew T M.

In: Journal of Cellular Physiology, Vol. 115, No. 3, 1983, p. 249-254.

Research output: Contribution to journalArticle

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AU - Crocker, J.

AU - Vaughan, Andrew T M

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N2 - Cells of the human promyelocytic HL60 line were induced to differentiate into granulocyte-like cells with dimethylsulphoxide (DMSO) or macrophage-like cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). The synthesis of Cathepsin B by these cells was studied by immunoperoxidase staining and assay of cell lysates using the fluorimetric substrate benzoyloxycarbonyl-phenylanalyl-arginine-4-methyl-7-coumarylamide. Only 2-5% of the uninduced HL60 cells and DMSO-induced cells were immunohistochemically positive for Cathepsin B, compared with over 80% of the TPA-induced cells. Cathepsin B activity was lowest in the lysates of uninduced HL60s. DMSO-induced cells contained 1.5-2-fold the enzyme activity of HL60s and TPA-induced cell lysates demonstrated 5-14-fold the activity of uninduced HL60s. Induction of Cathepsin B synthesis was therefore associated with differentiation of the promyelocytes into cells of the monocyte/macrophage type, but not granulocyte-like cells. Cathepsin B was located immunohistochemically in human palatine tonsils. The enzyme was only demonstrated within macrophages in these tissues. Cathepsin B may therefore be a useful immunohistochemical marker for malignant and nonmalignant cells of the monocyte/macrophage lineage.

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