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2 ', 3 '-dideoxycytidine represses thymidine kinases 1 and 2 expression in T-lymphoid cells

机译：2'，3'-二脱氧胞苷可抑制T淋巴细胞中的胸苷激酶1和2表达

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摘要

In vitro culture of H9 human lymphoid cells in the presence of 5.0 muM 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. (C) 2003 Elsevier Inc. All rights reserved. [References: 32]
机译：在存在5.0μM二脱氧胞苷（ddC）的情况下，体外培养H9人淋巴样细胞约40-45天，选择的细胞（H9-ddC细胞）对药物具有耐药性并对AZT（齐多夫定）具有交叉耐药性和5-氟-2'-脱氧尿苷（FdUR）。在这些细胞中对AZT和FdUR交叉耐药的主要机制是胸苷激酶（TK）的细胞活性低。为了探索H9-ddC细胞中TK活性降低的分子机制，进行了TK1和TK2的mRNA表达以及TK1蛋白的蛋白质印迹分析。 RT-PCR分析显示，在H9-ddC细胞中，TK1和TK2 mRNA的表达分别降低到27.1％和79.4％。通过在H9-ddC细胞的蛋白质印迹中不存在可检测到的TK1蛋白带，证实了TK1基因表达的降低。这些结果表明在ddC的存在下长期治疗H9细胞会下调TK1和TK2基因的表达，并降低耐药细胞中TK的表达和活性。 （C）2003 Elsevier Inc.保留所有权利。 [参考：32]

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