将胞苷脱氨酶互补DNA转入造血细胞。

Transfer of the cytidine deaminase cDNA into hematopoietic cells.

作者信息

Flasshove M, Frings W, Schröder J K, Moritz T, Schütte J, Seeber S

机构信息

Department of Internal Medicine (Cancer Research), West German Cancer Center, University of Essen Medical School.

出版信息

Leuk Res. 1999 Nov;23(11):1047-53. doi: 10.1016/s0145-2126(99)00128-9.

DOI:10.1016/s0145-2126(99)00128-9

PMID:10576510

Abstract

In order to investigate whether transfer of the cytidine deaminase (CDD) cDNA would increase chemotherapy resistance to cytosine arabinoside (ara-C) we used a retroviral vector expressing both, neomycin phosphotransferase and the CDD cDNA, to transduce hematopoietic cells from cell lines and from murine bone marrow (BM). After coculture on producer clones with a viral titer of 1 x 10(5) CFU/ml and up to 3-fold increased CDD enzymatic activity, WEHI-3 cell line and primary hematopoietic cells were exposed to ara-C in clonogenic assays. A transduction efficiency of 34.8 +/- 6.2% could be determined for BM clonogenic progenitor cells by G418 resistance. We could observe significantly more colonies (77 +/- 3.1%) surviving from transduced primary BM cells than from mock cells (51.7 +/- 9.3%) at 10(-8) mol/l ara-C. At 10(-7) mol/l ara-C 8.7% of BM cells became absolutely resistant after retroviral transduction. Our data confirm that CDD represents another candidate gene for increasing resistance to cytotoxic drugs in hematopoietic cells.

摘要

为了研究胞苷脱氨酶（CDD）cDNA的转移是否会增加对阿糖胞苷（ara-C）的化疗耐药性，我们使用了一种同时表达新霉素磷酸转移酶和CDD cDNA的逆转录病毒载体，来转导细胞系和小鼠骨髓（BM）中的造血细胞。在与病毒滴度为1×10⁵CFU/ml的生产克隆共培养后，CDD酶活性增加了3倍，在克隆形成试验中，将WEHI-3细胞系和原代造血细胞暴露于ara-C。通过G418抗性可确定BM克隆形成祖细胞的转导效率为34.8±6.2%。在10⁻⁸mol/l ara-C浓度下，我们观察到转导的原代BM细胞存活的集落（77±3.1%）明显多于mock细胞（51.7±9.3%）。在10⁻⁷mol/l ara-C浓度下，逆转录病毒转导后8.7%的BM细胞变得完全耐药。我们的数据证实，CDD是增加造血细胞对细胞毒性药物耐药性的另一个候选基因。