Efficacy of lentivirus-mediated Drosophila melanogaster deoxyribonucleoside kinase combined with (E)-5-(2-bromovinyl)-2'-deoxyuridine or 1-ß-D-arabinofuranosylthymine therapy in human keloid fibroblasts

Abstract

Keloid scarring is a type of fibroproliferative disease with a high recurrence rate. However, no effective treatment is currently available. Combined therapy with recombinant lentivirus-mediated Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) and prodrug has been widelystudiedandusedforcancertreatment.Duetothesimilarities between keloid scars and tumors, the aim of the present study was to investigate the efficacy of a Dm-dNK/nucleoside analog system for the treatment of keloid scars. Recombinant lentivirus expression of the Dm-dNK suicide gene was assessed. Western blotting was used to examine the protein expression of lentivirus mediated Dm-dNK in keloid fibroblasts. Enzyme activity assays were conducted using [3H]-labeled substrates. Furthermore, cytotoxicity and bystander effects were evaluated using MTT assays. The expression of green fluorescent protein was observed using fluorescence microscope and results indicated that there was no notable difference in lentivirus infectivity between the multiplicity of infection (MOI) of 1 and 10 in cells. Notably, western blotting revealed that Dm-dNK was stably expressed in keloid fibroblasts and the enzymatic activity assays revealed that the enzyme was activated following introduction into the keloid fibroblasts via the lentivirus. The cytotoxicity and bystander effects of Dm-dNK combined with cytotoxic nucleoside analogs were both observed in Dm-dNK+ keloid fibroblasts. These results demonstrated that the lentivirus-mediated Dm-dNK therapy may be effective in treating keloid fibroblasts, which provides some evidence for the use of Dm-dNK/prodrug therapy for keloid treatment in vivo in the future.