Properties and levels of deoxynucleoside kinases in normal and tumor cells; implications for chemotherapy

Staffan Eriksson, Elias Arnér, T. Spasokoukotskaja, Liya Wang, Anna Karlsson, Otte Brosjö, Peter Gunvén, Gunnar Julusson, Jan Liliemark

Research output: Contribution to journalArticle

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Abstract

Deoxynucleoside kinases are key enzymes in deoxyribonucleoside salvage, activating several clinically important chemotherapeutic drugs. The four known kinases, cytosolic thymidine kinase (TK1) and deoxycytidine kinase (dCK) and the mitochondrial thymidine kinase (TK2) and deoxyguanosine kinase (dGK), have been purified and characterized as to the subunit structure as well as specificity with a large number of analogs. These results are summarized and used to establish selective assays for the four enzymes in crude extracts of normal and malignant human peripheral blood mononuclear cells, gastrointestinal tissues and sarcomas. TK2 and dGK activities were found at low levels in all tissues, possibly correlated to the content of mitochondria. TK1 activity was detected only in samples containing a significant number of S phase cells. We have measured dCK activity as well as dCK polypeptide level by immuno blotting in these extracts. High levels of dCK were found in normal mononuclear leukocytes (91-145 ng dCK/mg protein) and in B-cell chronic lymphocytic leukemia (80 ± 30 ng/mg, n = 23). Hairy cell leukemia contained lower levels (28 ± 23 ng/mg, n = 7), as did unexpectedly three samples of T-cell chronic lymphocytic leukemia (18 ± 14 ng/mg). Phytohemaglutinine stimulation of normal lymphocytes did not lead to any substantial increase in either dCK activity or expression (less than 2.5-fold). In colon adenocarcinomas, the dCK content was significantly higher (21 ± 9.3 ng/mg, n = 20) than in normal colon mucosa (8.2 ± 3.7 ng/mg, n = 19, p <0.05). A similar pattern of dCK expression was found in gastric adenocarcinomas (21 ± 13 ng/mg, n = 5) and normal ventricular mucosa (6.2 ± 5.4 ng/mg, n = 5, p <0.15). One leiomyosarcoma and one extra-skeletal osteosarcoma showed a dCK levels comparable to those found in normal lymphocytes (84 ± 6 and 109 ± 4 ng/mg), while other sarcoma samples contained levels comparable to the gastrointestinal adenocarcinomas (20 ± 7 ng/mg, n = 12). We confirm that dCK is expressed constitutively and predominantly in lymphoid cells, but conclude that a significant expression may be found in non-lymphoid tissues as well, with increased levels in the corresponding tumor tissue. 2-Chlorodeoxyadenosine (CdA), an antileukemic agent used in treatment of hairy cell leukemia and chronic lymphocytic leukemias (B-CLL), is phosphorylated by dCK which was used as the selective substrate for this enzyme. A study was performed to investigate if there was a correlation between the dCK levels and the response to CdA treatment. In case of B-CLL, 21 patients that responded (complete and partial response) to CdA treatment, showed a significantly higher CdA phosphorylation than 13 patients not responding to CdA treatment (456 ± 170 pmole/mg/min versus 309 ± 97 pmole/mg/min, p <0.01). We conclude that the level of CdA phosphorylation is correlated to the response of leukemias to CdA treatment.

Original languageEnglish
Pages (from-to)13-25
Number of pages13
JournalAdvances in Enzyme Regulation
Volume34
Issue numberC
DOIs
Publication statusPublished - 1994

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Deoxycytidine Kinase
Chemotherapy
Tumors
Cladribine
Cells
Drug Therapy
Neoplasms
deoxyguanosine kinase
Tissue
Hairy Cell Leukemia
Phosphorylation
Adenocarcinoma
Lymphocytes
Thymidine Kinase
B-Cell Chronic Lymphocytic Leukemia
Sarcoma
deoxyribonucleoside kinases
Colon
Mucous Membrane
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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Properties and levels of deoxynucleoside kinases in normal and tumor cells; implications for chemotherapy. / Eriksson, Staffan; Arnér, Elias; Spasokoukotskaja, T.; Wang, Liya; Karlsson, Anna; Brosjö, Otte; Gunvén, Peter; Julusson, Gunnar; Liliemark, Jan.

In: Advances in Enzyme Regulation, Vol. 34, No. C, 1994, p. 13-25.

Research output: Contribution to journalArticle

Eriksson, S, Arnér, E, Spasokoukotskaja, T, Wang, L, Karlsson, A, Brosjö, O, Gunvén, P, Julusson, G & Liliemark, J 1994, 'Properties and levels of deoxynucleoside kinases in normal and tumor cells; implications for chemotherapy', Advances in Enzyme Regulation, vol. 34, no. C, pp. 13-25. https://doi.org/10.1016/0065-2571(94)90006-X
Eriksson, Staffan ; Arnér, Elias ; Spasokoukotskaja, T. ; Wang, Liya ; Karlsson, Anna ; Brosjö, Otte ; Gunvén, Peter ; Julusson, Gunnar ; Liliemark, Jan. / Properties and levels of deoxynucleoside kinases in normal and tumor cells; implications for chemotherapy. In: Advances in Enzyme Regulation. 1994 ; Vol. 34, No. C. pp. 13-25.
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T1 - Properties and levels of deoxynucleoside kinases in normal and tumor cells; implications for chemotherapy

AU - Eriksson, Staffan

AU - Arnér, Elias

AU - Spasokoukotskaja, T.

AU - Wang, Liya

AU - Karlsson, Anna

AU - Brosjö, Otte

AU - Gunvén, Peter

AU - Julusson, Gunnar

AU - Liliemark, Jan

PY - 1994

Y1 - 1994

N2 - Deoxynucleoside kinases are key enzymes in deoxyribonucleoside salvage, activating several clinically important chemotherapeutic drugs. The four known kinases, cytosolic thymidine kinase (TK1) and deoxycytidine kinase (dCK) and the mitochondrial thymidine kinase (TK2) and deoxyguanosine kinase (dGK), have been purified and characterized as to the subunit structure as well as specificity with a large number of analogs. These results are summarized and used to establish selective assays for the four enzymes in crude extracts of normal and malignant human peripheral blood mononuclear cells, gastrointestinal tissues and sarcomas. TK2 and dGK activities were found at low levels in all tissues, possibly correlated to the content of mitochondria. TK1 activity was detected only in samples containing a significant number of S phase cells. We have measured dCK activity as well as dCK polypeptide level by immuno blotting in these extracts. High levels of dCK were found in normal mononuclear leukocytes (91-145 ng dCK/mg protein) and in B-cell chronic lymphocytic leukemia (80 ± 30 ng/mg, n = 23). Hairy cell leukemia contained lower levels (28 ± 23 ng/mg, n = 7), as did unexpectedly three samples of T-cell chronic lymphocytic leukemia (18 ± 14 ng/mg). Phytohemaglutinine stimulation of normal lymphocytes did not lead to any substantial increase in either dCK activity or expression (less than 2.5-fold). In colon adenocarcinomas, the dCK content was significantly higher (21 ± 9.3 ng/mg, n = 20) than in normal colon mucosa (8.2 ± 3.7 ng/mg, n = 19, p <0.05). A similar pattern of dCK expression was found in gastric adenocarcinomas (21 ± 13 ng/mg, n = 5) and normal ventricular mucosa (6.2 ± 5.4 ng/mg, n = 5, p <0.15). One leiomyosarcoma and one extra-skeletal osteosarcoma showed a dCK levels comparable to those found in normal lymphocytes (84 ± 6 and 109 ± 4 ng/mg), while other sarcoma samples contained levels comparable to the gastrointestinal adenocarcinomas (20 ± 7 ng/mg, n = 12). We confirm that dCK is expressed constitutively and predominantly in lymphoid cells, but conclude that a significant expression may be found in non-lymphoid tissues as well, with increased levels in the corresponding tumor tissue. 2-Chlorodeoxyadenosine (CdA), an antileukemic agent used in treatment of hairy cell leukemia and chronic lymphocytic leukemias (B-CLL), is phosphorylated by dCK which was used as the selective substrate for this enzyme. A study was performed to investigate if there was a correlation between the dCK levels and the response to CdA treatment. In case of B-CLL, 21 patients that responded (complete and partial response) to CdA treatment, showed a significantly higher CdA phosphorylation than 13 patients not responding to CdA treatment (456 ± 170 pmole/mg/min versus 309 ± 97 pmole/mg/min, p <0.01). We conclude that the level of CdA phosphorylation is correlated to the response of leukemias to CdA treatment.

AB - Deoxynucleoside kinases are key enzymes in deoxyribonucleoside salvage, activating several clinically important chemotherapeutic drugs. The four known kinases, cytosolic thymidine kinase (TK1) and deoxycytidine kinase (dCK) and the mitochondrial thymidine kinase (TK2) and deoxyguanosine kinase (dGK), have been purified and characterized as to the subunit structure as well as specificity with a large number of analogs. These results are summarized and used to establish selective assays for the four enzymes in crude extracts of normal and malignant human peripheral blood mononuclear cells, gastrointestinal tissues and sarcomas. TK2 and dGK activities were found at low levels in all tissues, possibly correlated to the content of mitochondria. TK1 activity was detected only in samples containing a significant number of S phase cells. We have measured dCK activity as well as dCK polypeptide level by immuno blotting in these extracts. High levels of dCK were found in normal mononuclear leukocytes (91-145 ng dCK/mg protein) and in B-cell chronic lymphocytic leukemia (80 ± 30 ng/mg, n = 23). Hairy cell leukemia contained lower levels (28 ± 23 ng/mg, n = 7), as did unexpectedly three samples of T-cell chronic lymphocytic leukemia (18 ± 14 ng/mg). Phytohemaglutinine stimulation of normal lymphocytes did not lead to any substantial increase in either dCK activity or expression (less than 2.5-fold). In colon adenocarcinomas, the dCK content was significantly higher (21 ± 9.3 ng/mg, n = 20) than in normal colon mucosa (8.2 ± 3.7 ng/mg, n = 19, p <0.05). A similar pattern of dCK expression was found in gastric adenocarcinomas (21 ± 13 ng/mg, n = 5) and normal ventricular mucosa (6.2 ± 5.4 ng/mg, n = 5, p <0.15). One leiomyosarcoma and one extra-skeletal osteosarcoma showed a dCK levels comparable to those found in normal lymphocytes (84 ± 6 and 109 ± 4 ng/mg), while other sarcoma samples contained levels comparable to the gastrointestinal adenocarcinomas (20 ± 7 ng/mg, n = 12). We confirm that dCK is expressed constitutively and predominantly in lymphoid cells, but conclude that a significant expression may be found in non-lymphoid tissues as well, with increased levels in the corresponding tumor tissue. 2-Chlorodeoxyadenosine (CdA), an antileukemic agent used in treatment of hairy cell leukemia and chronic lymphocytic leukemias (B-CLL), is phosphorylated by dCK which was used as the selective substrate for this enzyme. A study was performed to investigate if there was a correlation between the dCK levels and the response to CdA treatment. In case of B-CLL, 21 patients that responded (complete and partial response) to CdA treatment, showed a significantly higher CdA phosphorylation than 13 patients not responding to CdA treatment (456 ± 170 pmole/mg/min versus 309 ± 97 pmole/mg/min, p <0.01). We conclude that the level of CdA phosphorylation is correlated to the response of leukemias to CdA treatment.

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