Retrovirus-mediated Drosophila melanogaster multisubstrate deoxyribonucleoside kinase gene therapy of gastric cancer cells in vitro and in vivo.
OBJECTIVE
The cytotoxicity of the Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (Dm-dNK)/nucleoside analog system to human gastric carci-noma cells was evaluated in vitro and in vivo for the development of novel gene therapy for gastric cancer.


METHODS
MGC-803 and BGC-823 cells were infected with retroviral vectors containing the Dm-dNK gene expressing Dm-dNK in its active form. [(3)H]-labeled substrates were used for enzyme activity assays and cytotoxicities were evaluated in vitro by MTT assay and annexin-V-FITC labeled FACS methods. Bystander effect assay was performed by MTT assay using a mixture of cells with and without expressed Dm-dNK. Anti-tumor efficacy was also examined in vivo.


RESULTS
Nucleoside analogs were significantly more cytotoxic to Dm-dNK-expressing cells than to their parental counterparts, and death was attributable to apoptosis. Implementation of the Dm-dNK/nucleoside analog system was associated with 'bystander' effects in vitro and suppressed the growth of MGC-803 tumors in mice.


CONCLUSIONS
The Dm-dNK/nucleoside analoguesystem possesses significant potential as suicide gene therapy for human gastric carcinoma.
Zhu
Zhi
Z
Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University, North Nanjing Street 155, Shenyang 110001, P.R.China.
Zhao
Lei
L
He
Anning
A
Huang
Baojun
B
Karlsson
Anna
A
Xu
Huimian
H
Zheng
Xinyu
X
D000818:Animals; D045744:Cell Line, Tumor; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis* / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013274:Stomach Neoplasms / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
D017853:Phosphotransferases (Alcohol Group Acceptor); C020018:deoxyribonucleoside kinases