Duesberg P, Stindl R, Hehlmann R
Department of Molecular and Cell Biology, Stanley Hall, University of California, Berkeley, CA 94720, USA.
Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11283-8. doi: 10.1073/pnas.201398998. Epub 2001 Sep 11.
Cancer cells and aneuploid cell lines can acquire resistance against multiple unrelated chemotherapeutic drugs that are over 3,000-fold those of normal levels and display spontaneous resistances up to 20-fold of normal levels. Two different mechanisms were proposed for this phenotype: (i) classical mutation of drug metabolizing genes or (ii) chromosome reassortments, catalyzed by cancer- and cell line-specific aneuploidy, which generate, via new gene dosage combinations, a plethora of cancer phenotypes, including drug resistance. To distinguish between these mechanisms, we have asked whether three mouse cell lines can become drug resistant, from which two or three genes have been deleted, and on which multidrug resistance is thought to depend: Mdr1a, Mdr1b, and Mrp1. Because all three lines could acquire multidrug resistance and were aneuploid, whereas diploid mouse cells could not, we conclude that aneuploid cells become drug resistant via specific chromosome assortments, independent of putative resistance genes. We have asked further whether aneuploid drug-resistant Chinese hamster cells revert spontaneously to drug sensitivity in the absence of cytotoxic drugs at the high rates that are typical of chromosome reassortments catalyzed by aneuploidy or at the very low or zero rates (i.e., deletion) of gene mutation. We found that four drug-resistant hamster cell lines reverted to drug sensitivity at rates of about 2-3% per generation, whereas two closely related lines remained resistant under our conditions. Thus, the karyotypic instability generated by aneuploidy emerges as the common source of the various levels of drug resistance of cancer cells: minor spontaneous resistances reflect accidental chromosome assortments, the high selected resistances reflect complex specific assortments, and multidrug resistance reflects new combinations of unselected genes located on the same chromosomes as selected genes.
癌细胞和非整倍体细胞系能够获得对多种不相关化疗药物的抗性,这些抗性水平比正常水平高3000倍以上,并且表现出高达正常水平20倍的自发抗性。针对这种表型提出了两种不同的机制:(i)药物代谢基因的经典突变,或(ii)由癌症和细胞系特异性非整倍体催化的染色体重排,通过新的基因剂量组合产生大量癌症表型,包括耐药性。为了区分这些机制,我们研究了三种小鼠细胞系是否会产生耐药性,其中两个或三个基因已被删除,并且认为多药耐药性取决于这些基因:Mdr1a、Mdr1b和Mrp1。由于所有这三种细胞系都能获得多药耐药性且为非整倍体,而二倍体小鼠细胞则不能,因此我们得出结论,非整倍体细胞通过特定的染色体重排产生耐药性,与假定的耐药基因无关。我们进一步研究了非整倍体耐药的中国仓鼠细胞在没有细胞毒性药物的情况下是否会以非整倍体催化的染色体重排典型的高频率自发恢复药物敏感性,或者以基因突变的非常低或零频率(即缺失)恢复。我们发现,四个耐药仓鼠细胞系以每代约2 - 3%的频率恢复药物敏感性,而两个密切相关的细胞系在我们的条件下仍保持耐药性。因此,非整倍体产生的核型不稳定性成为癌细胞各种耐药水平的共同来源:轻微的自发抗性反映偶然的染色体重排,高度选择的抗性反映复杂的特定重排,多药耐药性反映与选择基因位于同一条染色体上的未选择基因的新组合。