2′, 3′-Dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells

Tieran Han, Marilyn Fernandez, Malancha Sarkar, Ram P. Agarwal

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

In vitro culture of H9 human lymphoid cells in the presence of 5.0 μM dideoxycytidine (ddC), for about 40-45 days, selected cells (H9-ddC cells), which were resistant to the drug and cross-resistant to AZT (zidovudine) and 5-fluoro-2′-deoxyuridine (FdUR). The major mechanism of cross-resistance to AZT and FdUR in these cells was low cellular activity of thymidine kinase (TK). To explore molecular mechanisms of the reduced TK activity in H9-ddC cells, the mRNA expression of TK1 and TK2 and western blot analysis of TK1 protein were performed. RT-PCR analysis revealed that in H9-ddC cells the expression of both TK1 and TK2 mRNA was reduced to 27.1% and 79.4%, respectively. The reduced TK1 gene expression was confirmed by an absence of a detectable TK1 protein band in western blot of H9-ddC cells. These results demonstrate that long-term treatment of H9 cells in the presence of ddC down-regulated TK1 and TK2 gene expression and reduced the expression and activity of TK in the resistant cells.

Original languageEnglish
Pages (from-to)835-842
Number of pages8
JournalLife Sciences
Volume74
Issue number7
DOIs
StatePublished - Jan 2 2004
Externally publishedYes

Fingerprint

Zalcitabine
Lymphocytes
Thymidine Kinase
Gene expression
Messenger RNA
Zidovudine
Western Blotting
Gene Expression
thymidine kinase 1
thymidine kinase 2
Proteins
Pharmaceutical Preparations
Polymerase Chain Reaction

Keywords

  • Cross-resistance
  • Deoxycytidine kinase
  • Dideoxycytidine
  • Gene down regulation
  • Thymidine kinase 2
  • Thymidine kinase1

ASJC Scopus subject areas

  • Pharmacology

Cite this

2′, 3′-Dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells. / Han, Tieran; Fernandez, Marilyn; Sarkar, Malancha; Agarwal, Ram P.

In: Life Sciences, Vol. 74, No. 7, 02.01.2004, p. 835-842.

Research output: Contribution to journalArticle

Han, Tieran ; Fernandez, Marilyn ; Sarkar, Malancha ; Agarwal, Ram P. / 2′, 3′-Dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells. In: Life Sciences. 2004 ; Vol. 74, No. 7. pp. 835-842.
@article{7a946ea10cf54bcb95e7bd261ff3d29f,
title = "2′, 3′-Dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells",
abstract = "In vitro culture of H9 human lymphoid cells in the presence of 5.0 μM dideoxycytidine (ddC), for about 40-45 days, selected cells (H9-ddC cells), which were resistant to the drug and cross-resistant to AZT (zidovudine) and 5-fluoro-2′-deoxyuridine (FdUR). The major mechanism of cross-resistance to AZT and FdUR in these cells was low cellular activity of thymidine kinase (TK). To explore molecular mechanisms of the reduced TK activity in H9-ddC cells, the mRNA expression of TK1 and TK2 and western blot analysis of TK1 protein were performed. RT-PCR analysis revealed that in H9-ddC cells the expression of both TK1 and TK2 mRNA was reduced to 27.1{\%} and 79.4{\%}, respectively. The reduced TK1 gene expression was confirmed by an absence of a detectable TK1 protein band in western blot of H9-ddC cells. These results demonstrate that long-term treatment of H9 cells in the presence of ddC down-regulated TK1 and TK2 gene expression and reduced the expression and activity of TK in the resistant cells.",
keywords = "Cross-resistance, Deoxycytidine kinase, Dideoxycytidine, Gene down regulation, Thymidine kinase 2, Thymidine kinase1",
author = "Tieran Han and Marilyn Fernandez and Malancha Sarkar and Agarwal, {Ram P.}",
year = "2004",
month = "1",
day = "2",
doi = "10.1016/j.lfs.2003.07.023",
language = "English",
volume = "74",
pages = "835--842",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "7",

}

TY - JOUR

T1 - 2′, 3′-Dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells

AU - Han, Tieran

AU - Fernandez, Marilyn

AU - Sarkar, Malancha

AU - Agarwal, Ram P.

PY - 2004/1/2

Y1 - 2004/1/2

N2 - In vitro culture of H9 human lymphoid cells in the presence of 5.0 μM dideoxycytidine (ddC), for about 40-45 days, selected cells (H9-ddC cells), which were resistant to the drug and cross-resistant to AZT (zidovudine) and 5-fluoro-2′-deoxyuridine (FdUR). The major mechanism of cross-resistance to AZT and FdUR in these cells was low cellular activity of thymidine kinase (TK). To explore molecular mechanisms of the reduced TK activity in H9-ddC cells, the mRNA expression of TK1 and TK2 and western blot analysis of TK1 protein were performed. RT-PCR analysis revealed that in H9-ddC cells the expression of both TK1 and TK2 mRNA was reduced to 27.1% and 79.4%, respectively. The reduced TK1 gene expression was confirmed by an absence of a detectable TK1 protein band in western blot of H9-ddC cells. These results demonstrate that long-term treatment of H9 cells in the presence of ddC down-regulated TK1 and TK2 gene expression and reduced the expression and activity of TK in the resistant cells.

AB - In vitro culture of H9 human lymphoid cells in the presence of 5.0 μM dideoxycytidine (ddC), for about 40-45 days, selected cells (H9-ddC cells), which were resistant to the drug and cross-resistant to AZT (zidovudine) and 5-fluoro-2′-deoxyuridine (FdUR). The major mechanism of cross-resistance to AZT and FdUR in these cells was low cellular activity of thymidine kinase (TK). To explore molecular mechanisms of the reduced TK activity in H9-ddC cells, the mRNA expression of TK1 and TK2 and western blot analysis of TK1 protein were performed. RT-PCR analysis revealed that in H9-ddC cells the expression of both TK1 and TK2 mRNA was reduced to 27.1% and 79.4%, respectively. The reduced TK1 gene expression was confirmed by an absence of a detectable TK1 protein band in western blot of H9-ddC cells. These results demonstrate that long-term treatment of H9 cells in the presence of ddC down-regulated TK1 and TK2 gene expression and reduced the expression and activity of TK in the resistant cells.

KW - Cross-resistance

KW - Deoxycytidine kinase

KW - Dideoxycytidine

KW - Gene down regulation

KW - Thymidine kinase 2

KW - Thymidine kinase1

UR - http://www.scopus.com/inward/record.url?scp=0344196764&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0344196764&partnerID=8YFLogxK

U2 - 10.1016/j.lfs.2003.07.023

DO - 10.1016/j.lfs.2003.07.023

M3 - Article

VL - 74

SP - 835

EP - 842

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 7

ER -