Selective killing of glioma cells in culture and in vivo by retrovirus transfer of the herpes simplex virus thymidine kinase gene.
Selective killing of glioma cells in culture and in vivo by retrovirus transfer of the herpes simplex virus thymidine kinase gene.
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发表时间：
1991-06
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通讯作者：
Z. Ezzeddine;R. Martuza;D. Platika;M. Short;A. Malick;B. Choi;X. Breakefield
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作者：
Z. Ezzeddine;R. Martuza;D. Platika;M. Short;A. Malick;B. Choi;X. Breakefield
将单纯疱疹病毒1型（HSV-1）的胸苷激酶基因（tk）插入逆转录病毒载体中，在Moloney鼠白血病病毒长末端重复序列的增强子-启动子元件的转录控制下进行转录。复制缺陷型病毒颗粒通过将载体DNA转染到包装细胞系psi 2中获得，并用于感染培养中的C6大鼠神经胶质瘤衍生细胞系。这些细胞对核苷类似物更昔洛韦的毒性作用的敏感性被发现通过HSV-1 tk基因的转移而显著增加。感染和未感染细胞之间的敏感性差异定义了更昔洛韦浓度，该浓度可用于选择性地杀死基本上所有感染的细胞，而保留未感染的细胞。C6胶质瘤细胞皮下导入裸鼠体内具有高度致瘤性。腹腔注射更昔洛韦可抑制携带HSV-1 tk基因的C6衍生细胞而非亲代C6细胞产生的肿瘤生长。这项工作证明了由HSV-1 ks基因表达的胸苷激酶在使脑肿瘤细胞对核苷类似物的毒性作用敏感方面的有效性。因此，逆转录病毒载体应该证明在选择性地将这种杀伤基因传递到神经系统中的分裂肿瘤细胞中是有用的，其中大多数内源性细胞不分裂。
The thymidine kinase gene (tk) of herpes simplex type 1 virus (HSV-1) was inserted into a retroviral vector under the transcriptional control of the enhancer-promoter element of the Moloney murine leukemia virus long terminal repeat. Replication-defective viral particles were obtained by transfection of vector DNA into the packaging cell line psi2 and were used to infect C6 rat glioma-derived cell lines in culture. The sensitivity of these cells to the toxic effects of the nucleoside analog ganciclovir was found to be significantly increased by transfer of the HSV-1 tk gene. The difference in sensitivity between infected and uninfected cells defined ganciclovir concentrations that could be used to selectively kill essentially all infected cells while sparing uninfected ones. C6 glioma cells introduced subcutaneously into nude mice were highly tumorigenic. Growth of tumors produced from C6-derived cells bearing the HSV-1 tk gene, but not parental C6 cells, could be inhibited by intraperitoneal administration of ganciclovir. This work demonstrates the effectiveness of the thymidine kinase expressed by the HSV-1 ks gene in sensitizing brain tumor cells to the toxic effects of nucleoside analogs. Retrovirus vectors should thus prove useful in the selective delivery of this killer gene to dividing tumor cells in the nervous system, where most endogenous cells are not dividing.