Resistance to cytosine arabinoside by retrovirally mediated gene transfer of human cytidine deaminase into murine fibroblast and hematopoietic cells.
Resistance to cytosine arabinoside by retrovirally mediated gene transfer of human cytidine deaminase into murine fibroblast and hematopoietic cells.
复制标题
通过逆转录病毒介导的人胞苷脱氨酶基因转移至小鼠成纤维细胞和造血细胞中，对阿糖胞苷产生抗性。
作者：
R. Momparler;N. Eliopoulos;V. Bovenzi;S. Létourneau;M. Greenbaum;D. Cournoyer
胞嘧啶核苷类似物阿糖胞苷（ARA-C）产生的剂量限制性造血毒性是限制其用于治疗肿瘤性疾病的主要因素之一。一个有趣的方法来克服这个问题将是插入一个基因的耐药性ARA-C在正常的造血细胞，以保护他们免受药物毒性。ARA-C被胞苷脱氨酶脱氨基导致其抗氧化活性丧失。本研究的目的是确定人胞苷脱氨酶基因转移到小鼠成纤维细胞和造血细胞是否会赋予ARA-C耐药性。逆转录病毒介导的人胞苷脱氨酶基因转移到3 T3成纤维细胞中导致该基因的前病毒RNA的有效表达和胞质提取物中胞苷脱氨酶活性的增加。这些细胞显示出对ARA-C的显著抗性，如通过该药物对集落形成、细胞生长和DNA合成的影响所确定的。通过逆转录病毒载体将人胞苷脱氨酶基因转移到小鼠骨髓细胞中，在克隆形成试验中赋予了对ARA-C的高水平耐药性。这些研究表明，胞苷脱氨酶基因可用于癌症基因治疗，保护正常造血细胞免受ARA-C和相关的胞嘧啶核苷类似物的细胞毒性作用。
Dose-limiting hematopoietic toxicity produced by the cytosine nucleoside analogue cytosine arabinoside (ARA-C) is one of the major factors that limit its use in the treatment of neoplastic diseases. An interesting approach to overcome this problem would be to insert a gene for drug resistance to ARA-C in normal hematopoietic cells to protect them from drug toxicity. The deamination of ARA-C by cytidine deaminase results in a loss of its antineoplastic activity. The objective of this study was to determine if gene transfer of human cytidine deaminase into murine fibroblast and hematopoietic cells would confer drug resistance to ARA-C. Retrovirally mediated transfer of the human cytidine deaminase gene into 3T3 fibroblasts resulted in efficient expression of the proviral RNA for this gene and in increased cytidine deaminase activity in cytoplasmic extracts. These cells showed marked resistance to ARA-C as determined by the effects of this drug on colony formation, cell growth, and DNA synthesis. The transfer of the human cytidine deaminase gene into murine bone marrow cells by the retroviral vector conferred a high level of drug resistance to ARA-C in clonogenic assays. These studies indicate that the cytidine deaminase gene could be used in cancer gene therapy by protecting normal hematopoietic cells against the cytotoxic effects of ARA-C and related cytosine nucleoside analogues.