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Robotic Mammosphere Assay for High-Throughput Screening in Triple-Negative Breast Cancer

Artikel i vetenskaplig tidskrift
Författare Paul A. Fitzpatrick
Nina Akrap
Elin Söderberg
H. Harrison
G. J. Thomson
Göran Landberg
Publicerad i Slas Discovery
Volym 22
Nummer/häfte 7
Sidor 827-836
ISSN 2472-5552
Publiceringsår 2017
Publicerad vid Sahlgrenska Cancer Center
Sidor 827-836
Språk en
Länkar dx.doi.org/10.1177/2472555217692321
Ämnesord cancer and cancer drugs, high-content screening, stem cells, cell-based assays, stem-cells, identification, resistance, inhibitor, target, Biochemistry & Molecular Biology, Biotechnology & Applied Microbiology, Chemistry
Ämneskategorier Cancer och onkologi

Sammanfattning

In order to identify novel treatment principles specifically affecting cancer stem cells in triple-negative breast cancer, we have developed a high-throughput screening method based on the mammosphere and anoikis resistance assays allowing us to screen compounds using a functional readout. The assay was validated against manual protocols and through the use of positive controls, such as the response to hypoxia and treatment with the known cancer stem cell-targeting compound salinomycin. Manual and robotic procedures were compared and produced similar results in cell handling, cell cultures, and counting techniques, with no statistically significant difference produced from either method. The variance between samples processed manually versus robotically was no greater than 0.012, while Levene's test of significance was 0.2, indicating no significant difference between mammosphere data produced manually or robotically. Through the screening of 989 FDA-approved drugs and a follow-up screen assessing the antineoplastic subgroup, we have identified three therapeutic compounds with the ability to modulate the breast cancer stem cell fraction in the triple-negative breast cancer cell line MDA-MB-231, highlighting their potential usage as stem cell-specific adjuvant treatments.

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