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Caveolin-1 implicated as a pro-invasive gene in high-grade glioma cell models: implementation of a 3d spheroid matrix invasion assay

Moriconi, Chiara ORCID:, Palmieri, Valentina, Tornillo, Giusy, Fillmore, Helen, Pilkington, Geoff and Gumbleton, Mark ORCID: 2017. Caveolin-1 implicated as a pro-invasive gene in high-grade glioma cell models: implementation of a 3d spheroid matrix invasion assay. Neuro-Oncology 19 (S1) , i21-i22. 10.1093/neuonc/now293.080

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INTRODUCTION: The poor prognosis associated with Glioblastoma multiforme (GBM) is multifactorial but includes the capacity of residual tumour cells not removed by surgery and resistant to radio-/chemo-therapy undergoing diffuse invasion into the surrounding brain tissue. Caveolin-1 (Cav-1) is the major structural and functional component of caveolae. In a number of tumour types Cav-1 is recognised to participate in cytoskeletal rearrangement, integrin-mediated adhesion and/or matrix remodelling. We proposed Cav-1 serves to promote invasion of GBM cells. To investigate this we have employed in an in-vitro 3D cellular invasion assay. METHOD: The human GBM cell lines, UP007 and UP029 established from primary cultures of biopsy-derived brain tumours (University of Portsmouth), U-87 MG (ECACC) and U-373 MG (ECACC) were genetically modified to stably knock-out Cav-1 using a Lentiviral Cav-1 shRNA approach; corresponding stably transfected non-target (NT) shRNA cell lines were generated as controls. Neuropheres were formed and embedded within an extracellular matrix (Matrigel™). Over a two-/four-day period (depending on cell line) the migration of cells away from the neurophere core (CORE) was quantified by image capture and processing (Image J) using a custom-developed MatLab script for pixel density analysis indicative of the density of migrating cellular material. RESULTS: Cav-1 knockout resulted in significant (P<0.01) reductions (42–54%) in cell invasion (reduced area of invasion and cell density within this area) of UP007, UP029 and U-87 MG; reductions comparable in magnitude to that of exposure to high concentrations of the Src inhibitor – saracatinib (2 μM). The wild-type and control (NT) U-373 MG showed very little invasive capacity with the corresponding Cav-1 negative cells showing only a consistent trend (P>0.05) towards reduced invasion. Depending upon the cell line the Cav-1 knockdown also resulted in reduced size and cellular density of the neurosphere core (UP007 and UP029) indicative of reduced proliferation and/or cell survival capacity. CONCLUSION: Using an in-vitro 3D cellular invasion assay we have found Cav-1 expression in a series of three GBM cell lines to promote cellular invasion capacity. Ongoing studies are addressing signalling mechanisms and the influence of the microenvironment.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Subjects: R Medicine > RM Therapeutics. Pharmacology
Uncontrolled Keywords: signal transduction extracellular matrix integrins heart failure biopsy brain tumor chemotherapy regimen glioblastoma adhesions caveolae caveolins cell line cell survival cytoskeleton genes glioma radio communications surgical procedures, operative country of wales neoplasms surgery specialty residual tumor cancer research brain tissue pixel script
Publisher: Oxford University Press
ISSN: 1523-5866
Date of First Compliant Deposit: 15 June 2017
Last Modified: 05 Dec 2022 12:39

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