[Transfection of eucaryotic cells using cytochrome CYP450 2B6 gene: sensitivity to cyclophosphamide].

Voprosy meditsinskoi khimii Pub Date : 2002-09-01
S N Kolomeĭchuk, O Iu Abakumova, M A Maslov, E A Kalashnikova, N G Morozova, G A Serebrennikova, V I Shvets, N N Sokolov
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引用次数: 0

Abstract

Sensitivity of 293 human epithelial kidney cells transfected by human cytochrome CYP450 gene to cyclophosphamide was investigated. Transfection was carried out by plasmid DNA containing CYP2B6 gene complexed with cationic liposomes. Liposomes were prepared from mixture of cationic lipids and cholesterol at different molar ratios. Experimental protocol included the following steps: transfection of epithelial kidney cells by complexes of plasmid DNA-cationic liposomes, clone selection in the medium with antibiotic Geneticin G418, selected clone harvesting and their treatment by cyclophosphamide as following cytotoxicity evaluation. It was shown that addition of 0.25 mM of cyclophosphamide resulted in death of 40-45% transfected cell population.

[细胞色素CYP450 2B6基因转染真核细胞:对环磷酰胺的敏感性]。
研究了转染人细胞色素CYP450基因的293人上皮肾细胞对环磷酰胺的敏感性。采用含CYP2B6基因的质粒DNA与阳离子脂质体配合转染。将阳离子脂质与胆固醇按不同摩尔比混合制备脂质体。实验方案包括以下步骤:用质粒dna -阳离子脂质体复合物转染肾上皮细胞,在抗生素genticin G418培养基中选择克隆,选择克隆收获,并用环磷酰胺处理,进行细胞毒性评估。结果表明,加入0.25 mM的环磷酰胺可导致40-45%的转染细胞死亡。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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