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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.accessioned2020-06-30T12:12:54Z
dc.date.available2020-06-30T12:12:54Z
dc.date.issued2016-12-26
dc.identifier.citationMorrison E, Wai P, Leonidou A, et al. Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures. J Vis Exp. 2016;(118):54738. Published 2016 Dec 26. doi:10.3791/54738en_US
dc.identifier.issn1940-087X
dc.identifier.otherARTN e54738
dc.identifier.urihttps://qmro.qmul.ac.uk/xmlui/handle/123456789/65332
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 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.en_US
dc.description.sponsorshipOpen Access fees were supported by Nexcelom Bioscience, LLCen_US
dc.description.sponsorshipThis work was funded by Breast Cancer Now. RN is the recipient of a Breast Cancer Now Career Development Fellowship (2011MaySF01)en_US
dc.language.isoenen_US
dc.publisherJournal of Visualized Experimentsen_US
dc.relation.ispartofJOVE-JOURNAL OF VISUALIZED EXPERIMENTS
dc.rightsCC-BY
dc.rightsAttribution 3.0 United States*
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/us/*
dc.subjectCancer Researchen_US
dc.subjectFunctionalen_US
dc.subjectgenomicsen_US
dc.subjectsiRNAen_US
dc.subjectspheroiden_US
dc.subjecttumoren_US
dc.subjectmicroenvironmenten_US
dc.titleUtilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Culturesen_US
dc.typeArticleen_US
dc.rights.holder2016. The authors
dc.identifier.doi10.3791/54738
pubs.author-urlhttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000397846600027&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=612ae0d773dcbdba3046f6df545e9f6aen_US
pubs.issue118en_US
pubs.notesNot knownen_US
pubs.publication-statusPublisheden_US
rioxxterms.funderDefault funderen_US
rioxxterms.identifier.projectDefault projecten_US


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