Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures
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Publisher
DOI
10.3791/54738
Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Issue
ISSN
1940-087X
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Show full item recordAbstract
The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the
discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully
appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from preclinical models to clinical trials.
Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening.
These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show
that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing
the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of
phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm
the spatial impact of these dependencies using immunohistochemistry.
Authors
Morrison, E; Wai, P; Leonidou, A; Bland, P; Khalique, S; Farnie, G; Daley, F; Peck, B; Natrajan, RCollections
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