Synergistic combination of valproic acid and oncolytic parvovirus H‐1PV as a potential therapy against cervical and pancreatic carcinomas H‐1PV and VPA.

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Synergistic combination of valproic acid and oncolytic parvovirus H‐1PV as a potential therapy against cervical and pancreatic carcinomas H‐1PV and VPA synergize to induce cell death, oxidative stress and DNA damage in cervical and pancreatic derived cancer cell lines Source data is available for this figure in the Supporting Information. Analysis of cell lysis. Cancer cell lines were seeded into 96‐well plates and infected with H‐1PV at the indicated MOI (pfu/cell) in combination or not with VPA (1 mM). The maximum viral dose tested is shown in grey. Cell lysis was measured after 72 h by means of the LDH assay, as described under Materials and Methods Section. Columns show average cell lysis values with standard deviation bars. Results from a typical experiment performed in quadruplicate are shown. *p<0.05; **p<0.01; ***p<0.001 as calculated by two sample Welch t‐test and adjustment for multiple tests with the Bonferroni method. ROS content. Representative FACS‐plots of DCFH‐DA‐stained cancer cells, untreated (black) or treated with VPA alone (green), with H‐1PV alone (red) or with both H‐1PV and VPA (blue). FACS analysis of cervical and pancreatic cancer cells was performed, at 24 and 48 h post‐treatment respectively, as described in Materials and Methods Section. Every experiment was performed at least in triplicate and repeated at least twice. A minimum of 20,000 events was acquired. DNA damage. Western blot analysis of the levels of the DNA damage marker protein phosphorylated H2AX (γ‐H2AX) in cervical and pancreatic cancer cells left untreated or treated with VPA, H‐1PV, or both. 20 µg total cell lysate were used for analysis. Actin was used as a loading control. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO Mol Med, Volume: 5, Issue: 10, Pages: 1537-1555, First published: 17 September 2013, DOI: (10.1002/emmm.201302796)