Department of Basic Oncology, I.U. Oncology Institute, 34093, Capa, Istanbul, Turkey.
Mol Biol Rep. 2010 Oct;37(7):3541-5. doi: 10.1007/s11033-010-0001-x. Epub 2010 Feb 25.
Head and neck cancer is the sixth most common cancer in the world and one of the most lethal cancers. Microsatellite instability is an important characteristic of tumor cells and is observed both in presence and absence of mismatch repair gene mutations. The importance of microsatellite instability in head and neck cancer is not well established due to the lack of a consensus panel and selection of different markers, criteria and methodological variances. The main objective of this study was to investigate the performance of a consensus panel of microsatellite repeats by automated fragment analysis. Matched tumor and normal tissue samples from 99 patients were analyzed using five mononucleotide markers. Following PCR the amplified fragments were analyzed by capillary electrophoresis on an ABI 310 genetic analyzer. Microsatellite instability was observed in 26 patients. In 17 patients instability was detected at multiple loci. NR21 and BAT25 were the most frequently altered targets. These two mononucleotide markers could detect all samples displaying high-instability. In this study we describe a standardized fluorescent multiplex PCR combined with computerized analysis, which allows rapid and accurate analysis of a high number of samples and obviates the need to compare tumors with matching normal tissue.
头颈部癌症是世界上第六大常见癌症,也是最致命的癌症之一。微卫星不稳定性是肿瘤细胞的一个重要特征,在错配修复基因突变的存在和不存在的情况下都可以观察到。由于缺乏共识小组以及对不同标志物、标准和方法差异的选择,微卫星不稳定性在头颈部癌症中的重要性尚未得到充分确立。本研究的主要目的是通过自动化片段分析来研究微卫星重复的共识小组的性能。使用五个单核苷酸标记物分析了 99 名患者的匹配肿瘤和正常组织样本。在 PCR 之后,通过毛细管电泳在 ABI 310 遗传分析仪上分析扩增片段。在 26 名患者中观察到微卫星不稳定性。在 17 名患者中,在多个位点检测到不稳定性。NR21 和 BAT25 是最常改变的靶标。这两个单核苷酸标记物可以检测到所有显示高不稳定性的样本。在这项研究中,我们描述了一种标准化的荧光多重 PCR 结合计算机分析,它允许快速准确地分析大量样本,并且不需要将肿瘤与匹配的正常组织进行比较。