Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures.
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Embargo End Date
ICR Authors
Authors
Morrison, E
Wai, P
Leonidou, A
Bland, P
Khalique, S
Farnie, G
Daley, F
Peck, B
Natrajan, R
Wai, P
Leonidou, A
Bland, P
Khalique, S
Farnie, G
Daley, F
Peck, B
Natrajan, R
Document Type
Journal Article
Date
2016-12-26
Date Accepted
2016-11-03
Abstract
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 pre-clinical 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.
Citation
Journal of visualized experiments : JoVE, 2016, (118)
DOI
Source Title
Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
ISSN
1940-087X
eISSN
1940-087X
Research Team
Functional Genomics
Vannini Group
Vannini Group