Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures.
Date
2016-12-26Author
Morrison, E
Wai, P
Leonidou, A
Bland, P
Khalique, S
Farnie, G
Daley, F
Peck, B
Natrajan, R
Type
Journal Article
Metadata
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 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.
Subject
Cell Line, Tumor
Spheroids, Cellular
Humans
Neoplasms
Receptor, erbB-2
Antineoplastic Agents
Genomics
Phosphatidylinositol 3-Kinases
Class I Phosphatidylinositol 3-Kinases
Research team
Functional Genomics
Vannini Group
Language
eng
Date accepted
2016-11-03
License start date
2016-12-26
Citation
Journal of visualized experiments : JoVE, 2016, (118)
Publisher
JOURNAL OF VISUALIZED EXPERIMENTS