Zheng X, Johansson M, Karlsson A
Karolinska Institute, Division of Clinical Virology, Huddinge University Hospital, S-141 86 Stockholm, Sweden.
Biochem Biophys Res Commun. 2001 Nov 23;289(1):229-33. doi: 10.1006/bbrc.2001.5953.
We have recently shown that the overexpression of Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (Dm-dNK) in cancer cell lines increases the cells' sensitivity to several cytotoxic nucleoside analogs and the enzyme may accordingly be used as a suicide gene in combined gene/chemotherapy treatment of cancer. To further characterize the enzyme for possible use as a suicide gene, we constructed a replication-deficient retroviral vector that expressed either the wild-type enzyme that localizes to the cell nucleus or a mutant (arg247ser) that localizes to the cytosol. A thymidine kinase-deficient osteosarcoma cell line was transduced with the recombinant virus and we compared the sensitivity and bystander cell killing when the cell lines were incubated with the pyrimidine nucleoside analogs (E)-5-(2-bromovinyl)-2'-deoxyuridine and 1-beta-D-arabinofuranosylthymine. In summary, we showed that the cells' sensitivity and the efficiency of bystander cell killing were not dependent on whether Dm-dNK was located in the nucleus or cytosol.
我们最近发现，在癌细胞系中过表达果蝇多底物脱氧核糖核苷激酶（Dm-dNK）会增加细胞对几种细胞毒性核苷类似物的敏感性，因此该酶可作为联合基因/化疗治疗癌症的自杀基因。为了进一步鉴定该酶作为自杀基因的潜在用途，我们构建了一种复制缺陷型逆转录病毒载体，该载体表达定位于细胞核的野生型酶或定位于细胞质的突变体（arg247ser）。用重组病毒转导胸苷激酶缺陷型骨肉瘤细胞系，然后将这些细胞系与嘧啶核苷类似物（E）-5-（2-溴乙烯基）-2'-脱氧尿苷和1-β-D-阿拉伯呋喃糖基胸腺嘧啶一起孵育，比较其敏感性和旁观者细胞杀伤情况。总之，我们发现细胞的敏感性和旁观者细胞杀伤效率并不取决于Dm-dNK是位于细胞核还是细胞质中。
