Cancer gene therapy using a replication-competent herpes simplex virus type 1 vector.
Cancer gene therapy using a replication-competent herpes simplex virus type 1 vector.
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DOI：
10.1097/00000658-199809000-00009
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发表时间：
1998-09
影响因子：
9
通讯作者：
Sam S. Yoon;N. M. Carroll;E. Chiocca;Kenneth K. Tanabe
中科院分区：
文献类型：
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作者：
Sam S. Yoon;N. M. Carroll;E. Chiocca;Kenneth K. Tanabe
目的 作者研究了 hrR3（一种源自 1 型单纯疱疹病毒 (HSV 1) 的病毒载体）在体外和体内破坏结肠癌细胞的功效。还评估了添加前药更昔洛韦与 hrR3 感染相结合的效果。摘要背景数据大多数癌症基因治疗策略使用无法复制的病毒载体。 HSV 1载体hrR3能够复制，其复制对细胞具有细胞毒性。 hrR3 还拥有 HSV-胸苷激酶基因，该基因可将更昔洛韦转化为有毒代谢物。因此，向hrR3感染的细胞中添加更昔洛韦可能会增强hrR3破坏肿瘤细胞的能力。为了增加对肿瘤细胞的特异性，hrR3 具有突变的核糖核苷酸还原酶基因，并在具有高水平内源核糖核苷酸还原酶的细胞中选择性复制。活跃分裂的细胞（例如肿瘤细胞）具有高水平的内源性核糖核苷酸还原酶，用于合成 DNA 前体。作者对使用 HSV 1 载体治疗结直肠癌肝转移感兴趣。方法通过蛋白质印迹分析测定几种结肠癌细胞系和人肝细胞原代培养物中核糖核苷酸还原酶的表达。使用体外测定法测定了结肠癌细胞系中 hrR3 介导的细胞毒性。将人结肠癌细胞系HT29注射到裸鼠的胁腹中，然后瘤内注射hrR3。在添加和不添加腹膜内更昔洛韦的情况下评估肿瘤生长率。结果结肠癌细胞系中的核糖核苷酸还原酶水平远高于人肝细胞原代培养物中的水平。 hrR3 在体外有效破坏结肠癌细胞系。在HT29侧腹肿瘤中单次瘤内注射hrR3可显着降低肿瘤生长速度，并且施用更昔洛韦没有相加作用。结论 hrR3 复制的固有细胞毒性可在体外和体内有效地破坏结肠癌细胞。添加更昔洛韦不会在体内增强这种细胞毒性。未来，更有效和更具选择性的HSV 1载体可能有助于治疗癌症。
OBJECTIVE The authors investigate the efficacy of hrR3, a viral vector derived from herpes simplex virus type 1 (HSV 1), in destroying colon carcinoma cells in vitro and in vivo. The effect of adding the prodrug ganciclovir in combination with hrR3 infection also is assessed. SUMMARY BACKGROUND DATA Most cancer gene therapy strategies use viral vectors that are incapable of replication. The HSV 1 vector hrR3 is capable of replication, and its replication is cytotoxic to cells. hrR3 also possesses the HSV-thymidine kinase gene, which converts ganciclovir into a toxic metabolite. Thus, the addition of ganciclovir to hrR3-infected cells may enhance the ability of hrR3 to destroy tumor cells. To increase specificity for tumor cells, hrR3 has a mutated ribonucleotide reductase gene and replicates selectively in cells with high levels of endogenous rbonucleotide reductase. Actively dividing cells such as tumor cells have high levels of endogenous ribonucleotide reductase for synthesis of DNA precursors. The authors are interested in the use of HSV 1 vectors to treat liver metastases from colorectal cancer. METHODS Ribonucleotide reductase expression in several colon carcinoma cell lines and in primary cultures of human hepatocytes was determined by Western blot analysis. hrR3-mediated cytotoxicity in the colon carcinoma cell lines was determined using an in vitro assay. The human colon carcinoma cell line HT29 was injected into the flanks of nude mice followed by intratumoral injection of hrR3. Tumor growth rate was assessed with and without the addition of intraperitoneal ganciclovir. RESULTS Ribonucleotide reductase levels in colon carcinoma cell lines are much higher than in primary cultures of human hepatocytes. hrR3 efficiently destroys colon carcinoma cell lines in vitro. A single intratumoral injection of hrR3 into HT29 flank tumors significantly reduces tumor growth rate, and the administration of ganciclovir has no additive effect. CONCLUSIONS The inherent cytotoxicity of hrR3 replication effectively destroys colon carcinoma cells in vitro and in vivo. This cytotoxicity is not enhanced in vivo by the addition of ganciclovir. In the future, more efficacious and selective HSV 1 vectors may be useful in the treatment of cancer.