DNA amplification in multidrug, cross-resistant Chinese hamster ovary cells: molecular characterization and cytogenetic localization of the amplified DNA.
DNA amplification in multidrug, cross-resistant Chinese hamster ovary cells: molecular characterization and cytogenetic localization of the amplified DNA.
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DOI：
10.1083/jcb.103.4.1159
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发表时间：
1986-10
期刊：
影响因子：
--
通讯作者：
Kuo T
中科院分区：
文献类型：
--
作者：
Teeter LD;Atsumi S;Sen S;Kuo T
通过逐步增加长春新碱浓度的选择，建立了长春新碱耐药（VCR）中国仓鼠卵巢（CHO）细胞。这些细胞对许多结构或功能不相似的药物表现出多药交叉耐药性。耐药细胞的细胞遗传学分析显示存在双微体和扩大的染色体片段，从而暗示基因扩增作为一种可能的耐药机制。从其它多药交叉抗性CHO细胞系分离的扩增的DNA区段（Roninson，I. B.，H. T. Abelson，D. E. Housman，N.豪厄尔和A. Varshavsky，1984，Nature（Lond.），309：626-628）也在我们的VCR系中扩增。将该DNA片段用作探针，筛选VCR基因组DNA的粘粒文库，并检索重叠克隆。在VCR细胞中扩增所有这些片段，总计约45个酶（kb）。使用原位杂交，我们本地化的扩增结构域的长臂CHO染色体1或Z1。北方杂交分析表明，4.3 kb的mRNA由该扩增的DNA结构域编码，并在VCR细胞中过量产生。这些扩增的DNA片段在收购动物细胞中的多药交叉耐药性的参与的建议。
Vincristine-resistant (VCR) Chinese hamster ovary (CHO) cells have been established by stepwise selection in increasing concentrations of vincristine. These cells exhibit multidrug cross-resistance to a number of drugs that have no structural or functional similarities. Cytogenetic analyses of resistant cells revealed the presence of double minutes and expanded chromosomal segments, thus implicating gene amplification as a possible mechanism of resistance. An amplified DNA segment isolated from other multidrug cross-resistant CHO cell lines (Roninson, I. B., H. T. Abelson, D. E. Housman, N. Howell, and A. Varshavsky, 1984, Nature (Lond.), 309:626-628) is also amplified in our VCR lines. This DNA segment was used as a probe to screen a cosmid library of VCR genomic DNA, and overlapping clones were retrieved. All of these segments, totaling approximately 45 kilobases (kb), were amplified in VCR cells. Using in situ hybridization, we localized the amplification domain to the long arm of CHO chromosome 1 or Z1. Northern hybridization analysis revealed that a 4.3-kb mRNA was encoded by this amplified DNA domain and was over-produced in the VCR cells. Suggestions for the involvement of these amplified DNA segments in the acquisition of multidrug cross-resistance in animal cells are also presented.