Single-Cell Analysis of BRAF V600E and NRAS Q61R Mutation Status in Melanoma Cell Lines as Method Generation for Circulating Melanoma Cells.

Autor: Po JW; Centre for Circulating Tumour Cell Diagnostics & Research at the Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia. joseph.po@health.nsw.gov.au.; Centre for Oncology Education and Research Translation (CONCERT), Liverpool, NSW, Australia. joseph.po@health.nsw.gov.au.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia. joseph.po@health.nsw.gov.au., Ma Y; 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.; School of Medicine, University of New South Wales, Kensington, NSW, Australia., Luk AWS; Centre for Circulating Tumour Cell Diagnostics & Research at the Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia.; Charles Perkins Centre, University of Sydney, Camperdown, NSW, Australia., Lynch D; 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.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia., Balakrishnar B; Liverpool Hospital, Liverpool, NSW, Australia., Brungs D; 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.; Illawarra Cancer Centre, Wollongong Hospital, Wollongong, NSW, Australia., Azimi F; Liverpool Hospital, Liverpool, NSW, Australia., Cooper A; Centre for Oncology Education and Research Translation (CONCERT), Liverpool, NSW, Australia.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia.; Liverpool Hospital, Liverpool, NSW, Australia., Saricilar E; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia.; School of Medicine, University of New South Wales, Kensington, NSW, Australia.; Liverpool Hospital, Liverpool, NSW, Australia.; University of Sydney, Camperdown, NSW, Australia., Murthy V; 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.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia.; Liverpool Hospital, Liverpool, NSW, Australia., de Souza P; Centre for Circulating Tumour Cell Diagnostics & Research at the Ingham Institute for Applied Medical Research, Liverpool, NSW, Australia.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia.; School of Medicine, University of New South Wales, Kensington, NSW, Australia.; Liverpool Hospital, Liverpool, NSW, Australia.; School of Medicine, University of Wollongong, Wollongong, NSW, Australia., Becker TM; 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.; School of Medicine, Western Sydney University, Campbelltown, NSW, Australia.; School of Medicine, University of New South Wales, Kensington, NSW, Australia.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2265, pp. 277-286.
DOI: 10.1007/978-1-0716-1205-7_21
Abstrakt: 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 V600E /NRAS Q61R 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.
Databáze: MEDLINE