Evaluation of 5?-nucleotidase as biochemical marker in leukemias and lymphomas

Gutensohn, Wolf; Thiel, Eckhard; Emmerich, Bertold

doi:10.1007/bf01484440

Cited by 21 publications

(4 citation statements)

References 12 publications

(9 reference statements)

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“…CAT and ␤-Galactosidase Assays-Chloramphenicol acetyltransferase activity was measured at 37°C in a reaction medium containing 1 mM dibutyryl CoA (25 g/assay), 10 M D-threo[dichloroacetyl- [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C]chloramphenicol (56 mCi/mmol, 0.25 Ci/assay) in 0.25 M Tris, pH 8.0, buffer. The incubation was initiated by the addition of 2-50 l of cell extract in a total volume of 125 l and terminated after 1-6 h with the addition of 300 l of xylenes.…”

Section: Materials-fetalmentioning

confidence: 99%

“…In contrast, elevated activity of ecto-5Ј-NT has been found in breast carcinoma (11), gastric cancer (12), pancreatic cancer (13), chronic myelogenous leukemia (10,14), cutaneous T-cell lymphoma (15), and Walker 256 carcinoma (16). Human renal carcinoma cell lines induced to differentiate by butyric acid have demonstrated both an increase (Cur cell line) and a decrease (Caki cell line) in ecto-5Ј-NT activity (17), demonstrating a complex, and perhaps context-dependent, pattern of ecto-5Ј-NT expression in differentiation and malignant transformation.…”

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confidence: 95%

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Tissue-specific Regulation of the Ecto-5′-nucleotidase Promoter

Spychała

Zimmermann

Mitchell

1999

Journal of Biological Chemistry

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We have isolated the 5 region of the ecto-5-nucleotidase (low K m 5-NT) gene and established that a 969-base pair (bp) fragment confers cell-specific expression of a CAT reporter gene that correlates with the expression of endogenous ecto-5-NT mRNA and enzymatic activity. A 768-bp upstream negative regulatory region has been identified that conferred lymphocyte-specific negative regulation in a heterologous system with a 244-bp deoxycytidine kinase core promoter. DNase I footprinting identified several protected areas including Sp1, Sp1/ AP-2, and cAMP response element (CRE) binding sites within the 201-bp core promoter region and Sp1, NRE2a, TCF-1/LEF-1, and Sp1/NF-AT binding sites in the upstream regulatory region. Whereas the CRE site was essential in mediating the negative activity of the upstream regulatory region in Jurkat but not in HeLa cells, mutation of the Sp1/AP-2 site decreased promoter activity in both cell lines. Electrophoretic mobility shift assay analysis of proteins binding to the CRE site identified both ATF-1 and ATF-2 in Jurkat cells. Finally, phorbol 12-myristate 13-acetate increased the activity of both the core and the 969-bp promoter fragments, and this increase was abrogated by mutations at the CRE site. In summary, we have identified a tissue-specific regulatory region 5 of the ecto-5-NT core promoter that requires the presence of a functional CRE site within the basal promoter for its suppressive activity.

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Section: Materials-fetalmentioning

confidence: 99%

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confidence: 95%

Tissue-specific Regulation of the Ecto-5′-nucleotidase Promoter

Spychała

Zimmermann

Mitchell

1999

Journal of Biological Chemistry

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“…Consequently, under stress conditions, B-cells selectively release adenosine [11]. In addition, the activity of 5'-nucleotidase may have prognostic value in leukemias [12][13][14][15] and other diseases: it is decreased in the lymphocytes of immunodeficient patients with hypogammaglobulinemia [16][17][18] and patients with chronic lymphocytic leukemia [19], and in lymphoblasts of patients with acute T-cell lymphoblastic leukemia [12,20]. Low intracellular 5'-nucleotidase activity may contribute to the accumulation of toxic deoxyribonucleoside triphosphates in immunodeficient patients with adenosine deaminase deft-ciency, or in the presence of adenosine deaminase inhibitors and with purine nucleoside phosphorylase deficiency [21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Altered properties of human t-lymphoblast soluble low Km 5′-nucleotidase: Comparison with B-lymphoblast enzyme

Madrid‐Marina¹,

Lestan²,

Nowak³

et al. 1993

Leukemia Research

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Soluble low Km 5'-nucleotidases have been purified from human cultured T- and B-lymphoblasts to compare their properties and to examine the mechanism of different rates of nucleotide dephosphorylation. The enzyme from B-lymphoblasts (MGL-8) was 4385-fold purified with a specific activity of 114 mumol/min/mg, while the enzyme from T-lymphoblasts (CEM, MOLT-4) was 4355-fold purified with a specific activity of 35 mumol/min/mg. The activity of both enzymes have an absolute requirement for Mg++. The B-cell enzyme has maximum activity with Mg2+ > Mn2+ > Co2+, while the T-cell enzyme had maximum activity with Co2+ > Mn2+ > Mg2+. The optimum activity was at pH 7.4-9.0 for the B-cell enzyme and pH 9.0 for the T-cell enzyme. Substrate specificity was the same for both enzymes with the following relative Vmax values: CMP > UMP > dUMP > dCMP > dAMP > IMP > GMP > dIMP > dGMP. The Km values for AMP and IMP were 12 and 25 microM for the B-cell enzyme, and 7.0 and 12 microM for the T-cell enzyme. ATP and ADP are competitive inhibitors of these enzymes with apparent Ki values of 100 and 20 microM for the B-cell enzyme, and 44 microM and 8 microM for the T-cell enzyme, respectively. The apparent molecular mass by gel filtration column chromatography is 145 kD for the B-cell enzyme and 72 kDa for the T-cell enzyme. The subunit molecular masses by Western blots are 69.2 kD for both enzymes. These properties suggest that the B-lymphoblast enzyme is identical or similar to the enzyme from human placenta. However, the T-cell enzyme has some different properties. We conclude that these differences plus a lower content of low Km 5'-nucleotidase in T-cells may account for the decreased ability of T-lymphoblasts to dephosphorylate nucleotides and may contribute to the selective cytotoxicity of deoxyribonucleosides for T-lymphoblasts as compared to B-lymphoblasts.

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“…Among the acute leukemias, the lowest 5' NT levels were found in T and B forms and the highest in common blasts; in null and non-lymphoid blasts intermediate levels are usually present [2, 7-121. In the field of chronic lymphoid malignancies, many investigations of 5 ' NT distribution have been performed by biochemical means alone. These studies showed very low levels of enzyme activity in chronic B lymphocytic leukemia (B-CLL) cells [2, 8,. Furthermore 5' NT expression in lymphoproliferative disorders could be related to the hypothetical maturation stage reached by the disorder, as defined by immunological marker studies [17,18].…”

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confidence: 99%

5′ nucleotidase in chronic b cell leukemias: A cytochemical and ultrastructural study

et al. 1990

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We studied by cytochemical means the distribution of 5' nucleotidase (5' NT), a purine degradative enzyme, in the circulating lymphocytes of 24 healthy donors and 41 cases of chronic lymphoid leukemias, classified according to morphological and immunological criteria. About half the normal circulating lymphocytes were 5'NT positive and exhibited variable degrees of enzyme activity. Among chronic B lymphocytic leukemias we found high percentages of positive cells only in the phenotypically more mature cases. Moreover all cases of hairy cell, follicular cell, lymphoplasmacytic, and plasma cell leukemia showed moderate or weak 5' NT reactivity. Also one case of chronic T lymphocytic leukemia, CD8 positive, was moderately positive, while another, with large granular lymphocyte morphology, was completely negative. Electron microscopy revealed a discontinuous, granular plasma membrane reaction pattern, varying in intensity from case to case. In conclusion, our results confirm the usefulness of the 5' NT cytochemical reaction for identification of lymphoid populations at different stages of maturation in chronic B cell disorders.

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Evaluation of 5?-nucleotidase as biochemical marker in leukemias and lymphomas

Cited by 21 publications

References 12 publications

Tissue-specific Regulation of the Ecto-5′-nucleotidase Promoter

Tissue-specific Regulation of the Ecto-5′-nucleotidase Promoter

Altered properties of human t-lymphoblast soluble low Km 5′-nucleotidase: Comparison with B-lymphoblast enzyme

5′ nucleotidase in chronic b cell leukemias: A cytochemical and ultrastructural study

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