Schmitt Christina, Schulz Anna-Alice, Winkelmann Ria, Smith Kevin, Wild Peter J, Demes Melanie
Dr. Senckenberg Institute of Pathology, University Hospital Frankfurt, Frankfurt am Main 60590, Germany.
Wildlab, University Hospital Frankfurt MVZ GmbH, Frankfurt am Main 60590, Germany.
Oncotarget. 2021 Oct 26;12(22):2273-2282. doi: 10.18632/oncotarget.28092.
gene alterations are known to be involved in acquired resistance to epidermal growth factor receptor inhibition. amplifications present a potential therapeutic target in non-small cell lung cancer. Although next-generation sequencing (NGS) and fluorescence hybridization (FISH) are conventionally used to assess amplifications, there are currently no clinically defined cut-off values for NGS, with FISH still being the gold standard. A collective of 20 formalin-fixed paraffin-embedded lung cancer tissue samples (mean age 64 years) were selected based on increased gene copy number (CNV) status or the presence of mutations detected by NGS (GeneReader, QIAGEN) and were further assessed by FISH (/CEN7, Zytomed). Of these, 17 tumor samples were -amplified and one patient was found to have a rearrangement by NGS, while two samples had no gene alteration. In contrast to the NGS result, FISH analysis showed only one highly amplified sample and 19 negative samples. The single highly amplified case detected by FISH was also positive by NGS with a fold change (FC) of 3.18 and a mean copy number (CN) of 20.5. Therefore, for the assessment of amplifications using the QIAGEN NGS workflow, we suggest detecting amplified cases with an FC value of ≥ 3.0 and a CN value of ≥ 20.0 by FISH. In summary, NGS allows for DNA- and RNA-based analysis of specific gene amplifications, point mutations or rearrangements.
已知基因改变与表皮生长因子受体抑制的获得性耐药有关。扩增在非小细胞肺癌中是一个潜在的治疗靶点。虽然下一代测序（NGS）和荧光原位杂交（FISH）传统上用于评估扩增情况，但目前NGS尚无临床定义的临界值，FISH仍是金标准。基于基因拷贝数（CNV）状态增加或通过NGS（GeneReader，QIAGEN）检测到的突变，选取了20个福尔马林固定石蜡包埋的肺癌组织样本（平均年龄64岁），并通过FISH（/CEN7，Zytomed）进一步评估。其中，17个肿瘤样本有扩增，1例患者通过NGS发现有重排，2个样本无基因改变。与NGS结果相反，FISH分析仅显示1个高度扩增样本和19个阴性样本。FISH检测到的单个高度扩增病例通过NGS也呈阳性，倍数变化（FC）为3.18，平均拷贝数（CN）为20.5。因此，对于使用QIAGEN NGS工作流程评估扩增情况，我们建议通过FISH检测FC值≥3.0且CN值≥20.0的扩增病例。总之，NGS允许对特定基因扩增、点突变或重排进行基于DNA和RNA的分析。
