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dc.contributor.authorMorrison, E
dc.contributor.authorWai, P
dc.contributor.authorLeonidou, A
dc.contributor.authorBland, P
dc.contributor.authorKhalique, S
dc.contributor.authorFarnie, G
dc.contributor.authorDaley, F
dc.contributor.authorPeck, B
dc.contributor.authorNatrajan, R
dc.date.accessioned2017-03-24T14:48:44Z
dc.date.issued2016-12-26
dc.identifier.citationJournal of visualized experiments : JoVE, 2016, (118)
dc.identifier.issn1940-087X
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/516
dc.identifier.eissn1940-087X
dc.identifier.doi10.3791/54738
dc.description.abstractThe 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.
dc.formatElectronic
dc.languageeng
dc.language.isoeng
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.subjectCell Line, Tumor
dc.subjectSpheroids, Cellular
dc.subjectHumans
dc.subjectNeoplasms
dc.subjectReceptor, erbB-2
dc.subjectAntineoplastic Agents
dc.subjectGenomics
dc.subjectPhosphatidylinositol 3-Kinases
dc.subjectClass I Phosphatidylinositol 3-Kinases
dc.titleUtilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures.
dc.typeJournal Article
dcterms.dateAccepted2016-11-03
rioxxterms.versionofrecord10.3791/54738
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0
rioxxterms.licenseref.startdate2016-12-26
rioxxterms.typeJournal Article/Review
dc.relation.isPartOfJournal of visualized experiments : JoVE
pubs.issue118
pubs.notesNo embargo
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research/Functional Genomics
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology/Functional Genomics
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology/Vannini Group
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research/Functional Genomics
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology/Functional Genomics
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology/Vannini Group
pubs.publication-statusPublished
pubs.embargo.termsNo embargo
icr.researchteamFunctional Genomicsen_US
icr.researchteamVannini Groupen_US
dc.contributor.icrauthorPeck, Barrieen
dc.contributor.icrauthorKhalique, Sairaen
dc.contributor.icrauthorNatrajan, Rachaelen
dc.contributor.icrauthorLeonidou, Andrien


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https://creativecommons.org/licenses/by/4.0
Except where otherwise noted, this item's license is described as https://creativecommons.org/licenses/by/4.0