Myeloperoxidase: expression and modulation in a large panel of human leukemia-lymphoma cell lines
| Autor: | ZB Hu, W Ma, CC Uphoff, K Metge, SM Gignac, HG Drexler |
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| Rok vydání: | 1993 |
| Předmět: |
Base Sequence
Lymphoma Histocytochemistry Immunology Molecular Sequence Data Fluorescent Antibody Technique Gene Expression Cell Biology Hematology Blotting Northern Biochemistry Polymerase Chain Reaction Leukemia Lymphoid Tumor Cells Cultured Humans Tetradecanoylphorbol Acetate RNA Messenger Half-Life Peroxidase |
| Zdroj: | Blood. 82(5) |
| ISSN: | 0006-4971 |
| Popis: | Myeloperoxidase (MPO) is found exclusively in the azurophilic granules (primary lysosomes) of normal myelomonocytic cells. Cytochemical staining for MPO activity is used clinically to distinguish myeloid from lymphoid leukemias. We studied the expression of MPO at the RNA and protein level in 140 continuous human leukemia-lymphoma cell lines using classical cytochemistry, immunofluorescent staining with a specific monoclonal antibody, Northern blot analysis, and a reverse transcription-polymerase chain reaction (RT-PCR) amplification assay. Seventy-eight lymphoid leukemia, myeloma, and lymphoma cell lines were negative; only 3 pre-B-acute lymphoblastic leukemia (ALL) cell lines were MPO-positive. Two of these MPO-positive pre-B-ALL cell lines showed a trace expression after RT-PCR and Southern blotting corresponding to 4% to 6% of the transcripts found in other positive myeloid cell lines. The third pre-B-ALL cell line was positive in Northern blots and cytochemical/immunofluorescent staining; however, only few cells were weakly positive in the latter assay. Although 15 of 59 cell lines assigned to the myeloid, monocytic, megakaryocytic, or erythroid lineages were MPO-positive in Northern blots, those 15 and 13 additional cell lines showed bands of mRNA after RT-PCR. MPO protein was detected in all 16 Northern-positive cell lines; on the other hand, there were 4 cell lines that were protein-positive, but Northern- negative. Differentiation induced by protein kinase C activators 12-O- tetradecanoylphorbol 13-acetate and Bryostatin 1 or by all-trans retinoic acid was associated with a decrease in MPO mRNA in all 7 initially positive cell lines studied, even leading to the complete absence of transcripts, but the enzymatic activity of the differentiated cells was only slightly less than that of unstimulated cells. MPO expression could not be induced in 10 initially negative cell lines. The half-life of MPO mRNA was found to be about 6 hours and was not shortened by prior exposure of the cells to the differentiation- inducing agents. These results confirm that MPO expression is mainly associated with myelomonocytic cells, but also underline the notion that MPO cannot be used as an absolutely lineage-specific marker for the distinction of leukemic cells. MPO can be used as an excellent parameter to characterize the various stages of normal and induced differentiation. |
| Databáze: | OpenAIRE |
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