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通过DNA介导的基因转移对中国仓鼠卵巢细胞中甲氨蝶呤摄取缺陷进行互补。

Complementation of a methotrexate uptake defect in Chinese hamster ovary cells by DNA-mediated gene transfer.

作者信息

Underhill T M, Flintoff W F

机构信息

Department of Microbiology and Immunology, University of Western Ontario, London, Canada.

出版信息

Mol Cell Biol. 1989 Apr;9(4):1754-8. doi: 10.1128/mcb.9.4.1754-1758.1989.

DOI:10.1128/mcb.9.4.1754-1758.1989
PMID:2725523
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC362594/
Abstract

A methotrexate-resistant Chinese hamster ovary cell line deficient in methotrexate uptake has been complemented to methotrexate sensitivity by transfection with DNA isolated from either wild-type Chinese hamster ovary or human G2 cells. Primary and secondary transfectants regained the ability to take up methotrexate in a manner similar to that of wild-type cells, and in the case of those transfected with human DNA, to contain human-specific DNA sequences. The complementation by DNA-mediated gene transfer of this methotrexate-resistant phenotype provides a basis for the cloning of a gene involved in methotrexate uptake.

摘要

一种对甲氨蝶呤耐药且缺乏甲氨蝶呤摄取能力的中国仓鼠卵巢细胞系，通过用从野生型中国仓鼠卵巢细胞或人G2细胞中分离的DNA进行转染，恢复了对甲氨蝶呤的敏感性。原代和二代转染细胞重新获得了以类似于野生型细胞的方式摄取甲氨蝶呤的能力，对于那些用人类DNA转染的细胞，则含有人类特异性DNA序列。这种由甲氨蝶呤耐药表型的DNA介导基因转移所实现的互补作用，为克隆参与甲氨蝶呤摄取的基因提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80e8/362594/8bf8466b9b22/molcellb00052-0384-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80e8/362594/8bf8466b9b22/molcellb00052-0384-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80e8/362594/8bf8466b9b22/molcellb00052-0384-a.jpg

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Complementation of a methotrexate uptake defect in Chinese hamster ovary cells by DNA-mediated gene transfer.通过DNA介导的基因转移对中国仓鼠卵巢细胞中甲氨蝶呤摄取缺陷进行互补。
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Stable transfectants of human MCF-7 breast cancer cells with increased levels of the human folate receptor exhibit an increased sensitivity to antifolates.人叶酸受体水平升高的人MCF-7乳腺癌细胞稳定转染体对抗叶酸药物表现出更高的敏感性。
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本文引用的文献

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Isolation of transforming DNA: cloning the hamster aprt gene.转化DNA的分离：仓鼠aprt基因的克隆
Cell. 1980 Dec;22(3):817-23. doi: 10.1016/0092-8674(80)90558-9.
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Relationships between carrier-mediated transport of folate compounds by L1210 leukemia cells: evidence for multiplicity of entry routes with different kinetic properties expressed in plasma membrane vesicles.L1210白血病细胞对叶酸化合物的载体介导转运关系：质膜囊泡中表现出具有不同动力学特性的多种进入途径的证据。
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Transport of methotrexate in Chinese hamster ovary cells: a mutant defective in methotrexate uptake and cell binding.
甲氨蝶呤在中国仓鼠卵巢细胞中的转运：一种甲氨蝶呤摄取和细胞结合缺陷的突变体。
Arch Biochem Biophys. 1983 Jun;223(2):433-40. doi: 10.1016/0003-9861(83)90607-0.
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Affinity labeling of the 5-methyltetrahydrofolate/methotrexate transport protein of L1210 cells by treatment with an N-hydroxysuccinimide ester of [3H]methotrexate.通过用[3H]甲氨蝶呤的N-羟基琥珀酰亚胺酯处理对L1210细胞的5-甲基四氢叶酸/甲氨蝶呤转运蛋白进行亲和标记。
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Biochim Biophys Acta. 1983 Oct 26;735(1):123-30. doi: 10.1016/0005-2736(83)90267-5.
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Molecular cloning of a human DNA repair gene.一个人类DNA修复基因的分子克隆
Nature. 1984;310(5976):425-9. doi: 10.1038/310425a0.
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Isolation and characterization of a folate receptor from human placenta.人胎盘叶酸受体的分离与鉴定
J Biol Chem. 1981 Sep 25;256(18):9684-92.
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Isolation of high-molecular-weight DNA from mammalian cells.从哺乳动物细胞中分离高分子量DNA。
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Transport of folate compounds in L1210 cells: kinetic evidence that folate influx proceeds via the high-affinity transport system for 5-methyltetrahydrofolate and methotrexate.叶酸化合物在L1210细胞中的转运：动力学证据表明叶酸内流通过5-甲基四氢叶酸和甲氨蝶呤的高亲和力转运系统进行。
Cancer Res. 1986 Apr;46(4 Pt 1):1639-43.