Centre for Circulating Tumour Cell Diagnostics & Research at the Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia.
Centre for Oncology Education and Research Translation (CONCERT), Liverpool, NSW, Australia.
Methods Mol Biol. 2021;2265:277-286. doi: 10.1007/978-1-0716-1205-7_21.
Molecular testing of tumor biopsies allows for the identification of the key mutations driving a patient's cancer. However, this is limited to singular nodes and may not accurately reflect cancer heterogeneity. Circulating tumor cell (CTC) analyses offer a noninvasive method of interrogating the genomic profile of patient-derived tumor material. To date, molecular analysis of CTCs has relied on the characterization of bulk or pooled CTC lysates, limiting the detection of minor tumorigenic CTC subclones. Here, we show a workflow enabling BRAF/NRAS mutation detection from single cultured melanoma cells by combining micromanipulation and genomic material amplification methods. This workflow can be directly integrated into circulating tumor cell analysis applications.
肿瘤活检的分子检测可识别驱动患者癌症的关键突变。然而，这仅限于单一节点，并且可能无法准确反映癌症异质性。循环肿瘤细胞（CTC）分析提供了一种非侵入性的方法来研究患者来源的肿瘤材料的基因组特征。迄今为止，CTC 的分子分析依赖于对批量或汇集的 CTC 裂解物的特征描述，从而限制了对次要致瘤性 CTC 亚克隆的检测。在这里，我们展示了一种通过结合微操作和基因组物质扩增方法从单个培养的黑色素瘤细胞中检测 BRAF/NRAS 突变的工作流程。该工作流程可直接集成到循环肿瘤细胞分析应用中。
