tailieunhanh - Báo cáo khoa học: Altered deoxyribonucleotide pools in T-lymphoblastoid cells expressing the multisubstrate nucleoside kinase of Drosophila melanogaster

The multisubstrate nucleoside kinase of Drosophila melanogaster(Dm-dNK) can be expressed in human solid tumor cells and its unique enzymatic properties makes this enzyme a suicide gene candidate. In the present study,Dm-dNK was stably expressed in the CCRF-CEM and H9 T-lymphoblastoid cell lines. The expressed enzyme was localized to the cell nucleus and the enzyme retained its activity. | iFEBS Journal Altered deoxyribonucleotide pools in T-lymphoblastoid cells expressing the multisubstrate nucleoside kinase of Drosophila melanogaster Ada Bertoli1 Maribel Franco1 Jan Balzarini2 Magnus Johansson1 and Anna Karlsson1 1 Karolinska Institute Department of Laboratory Medicine Karolinska University Hospital Huddinge Stockholm Sweden 2 Rega Institute for MedicalResearch Leuven Belgium Keywords deoxyribonucleotide pools Dm-dNK nucleoside analogs suicide gene T-lymphoblastoid cell lines Correspondence A. Karlsson Karolinska Institute Department of Laboratory Medicine Karolinska University Hospital Huddinge S-141 86 Stockholm Sweden Fax 46 8 58587933 Tel 46 8 58587932 E-mail Present address Department of Experimental Medicine and BiochemicalSciences University of Rome Tor Vergata Rome I-00133 Italy Received 22 March 2005 revised 2 June 2005 accepted 7 June 2005 doi The multisubstrate nucleoside kinase of Drosophila melanogaster Dm-dNK can be expressed in human solid tumor cells and its unique enzymatic properties makes this enzyme a suicide gene candidate. In the present study Dm-dNK was stably expressed in the CCRF-CEM and H9 T-lymphoblastoid cell lines. The expressed enzyme was localized to the cell nucleus and the enzyme retained its activity. The Dm-dNK overexpressing cells showed w 200-fold increased sensitivity to the cytostatic activity of several nucleoside analogs such as the pyrimidine nucleoside analogs E -5- 2-bromovinyl -2 -deoxyuridine BVDU and 1-b-D-arabinofuranosyl-thymine araT but not to the antiherpetic purine nucleoside analogs ganciclovir acyclovir and penciclovir which may allow this technology to be applied in donor T cells and or rescue graft vs. host disease to permit modulation of alloreactivity after transplantation. The most pronounced effect on the steady-state dNTP levels was a two- to 10-fold increased dTTP pool in Dm-dNK expressing cells that were grown in the presence of 1