Grottke C, Mantwill K, Dietel M, Schadendorf D, Lage H
Institute of Pathology, Charité, Campus Mitte, Humboldt University Berlin, Berlin, Germany.
Int J Cancer. 2000 Nov 15;88(4):535-46. doi: 10.1002/1097-0215(20001115)88:4<535::aid-ijc4>3.0.co;2-v.
Malignant melanoma displays strong resistance against various antineoplastic drugs. The mechanisms conferring this intrinsic resistance are unclear. To better understand the molecular events associated with drug resistance in melanoma, a panel of human melanoma cell variants exhibiting low and high levels of resistance to 4 commonly used drugs in melanoma treatment, i.e., vindesine, etoposide, fotemustine and cisplatin, was characterized by differential display reverse transcription-polymerase chain reaction (DDRT-PCR). Of 269 mRNA fragments found to be altered in expression level by DDRT-PCR, a total of 11 cDNA clones was characterized after confirmation of a differential expression pattern by Northern blot analyses. These clones include 3 genes (DSM-1, DSM-3 and DSM-5) of known function, 4 previously sequenced genes (DSM-2, DSM-4, DSM-6 and DSM-7) of uncharacterized function and 4 novel genes (DSM-8-DSM-11) without match in GenBank. All of these genes exhibited altered mRNA expression in high level etoposide-resistant cells, whereby 7 genes (DSM-1-DSM-6 and DSM-8) were found to be decreased in the transcription rate in these etoposide-resistant cells. The mRNA synthesis of the remaining genes (DSM-7 and DSM-9-DSM11) was enhanced in high level etoposide-resistant melanoma cells. The expression of 5 (DSM-5 and DSM-7-DSM-10) of the cloned cDNA encoding mRNAs was modulated in various independently established drug-resistant melanoma cells, indicating to be associated with drug resistance. Further characterization of these genes may yield inside into the biology and development of drug resistance in malignant melanoma.
恶性黑色素瘤对多种抗肿瘤药物表现出强烈抗性。赋予这种内在抗性的机制尚不清楚。为了更好地理解黑色素瘤中与耐药性相关的分子事件，通过差异显示逆转录 - 聚合酶链反应（DDRT - PCR）对一组对黑色素瘤治疗中4种常用药物（长春地辛、依托泊苷、福莫司汀和顺铂）表现出低水平和高水平抗性的人黑色素瘤细胞变体进行了表征。在通过DDRT - PCR发现269个mRNA片段的表达水平发生改变后，通过Northern印迹分析确认差异表达模式后，对总共11个cDNA克隆进行了表征。这些克隆包括3个已知功能的基因（DSM - 1、DSM - 3和DSM - 5）、4个功能未明确的先前测序基因（DSM - 2、DSM - 4、DSM - 6和DSM - 7）以及4个在GenBank中无匹配项的新基因（DSM - 8 - DSM - 11）。所有这些基因在高水平依托泊苷抗性细胞中均表现出mRNA表达改变，其中7个基因（DSM - 1 - DSM - 6和DSM - 8）在这些依托泊苷抗性细胞中的转录速率降低。其余基因（DSM - 7和DSM - 9 - DSM11）的mRNA合成在高水平依托泊苷抗性黑色素瘤细胞中增强。编码mRNA的5个克隆cDNA（DSM - 5和DSM - 7 - DSM - 10）的表达在各种独立建立的耐药黑色素瘤细胞中受到调节，表明与耐药性相关。对这些基因的进一步表征可能有助于深入了解恶性黑色素瘤的生物学特性和耐药性的发展。
