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

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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/VPA co‐treatment leads to complete remission of established HeLa tumours Source data is available for this figure in the Supporting Information. Analysis of tumour growth. Animals (n = 8 per group) were treated with either PBS (control), VPA or a combination of both agents as described in Materials and Methods Section. Other animal groups are presented in Supporting Information Fig S7. The data shown represent average tumour volumes with standard deviation bars. Graphical summary of Kaplan–Meier survival analysis of tumour‐bearing nude rats treated as indicated. Animals were sacrificed when the tumour reached the maximum tolerable size of 4000 mm3 according to law. The difference between H‐1PV/VPA co‐treatment and treatment with H‐1PV alone was statistically significant (p = 0.0002 as calculated with the two‐sided log‐rank test and adjusted for multiple testing with the Bonferroni method). Other results from the statistical analysis are shown in Supporting Information Table S2. H‐1PV/VPA co‐treatment induces DNA damage and apoptosis in vivo. Two rats of each treatment group were sacrificed on day 19. Tumours were fixed, sectioned and examined morphologically either by H&E staining or by immunochemistry using antibodies against NS1, VP capsid proteins, γ‐H2AX (a DNA damage marker), or active cleaved caspase‐3 (an apoptosis marker). Scale bar = 50 µm. Higher levels of NS1, VP1 and VP2 viral proteins in tumour samples from VPA co‐treated animals. Tumour‐bearing animals (n = 3) treated with H‐1PV alone (total virus dose of 3.6 × 109 Vg/animal corresponding to 1.5 × 108 pfu/animals subdivided into two subdoses given at days 0 and 7) or co‐treated with H‐1PV and VPA (100 mg/kg) were sacrificed at day 14. Complete tumours were resected, homogenised and lysed. 20 µg of total cellular extracts were analysed by SDS–PAGE for the presence of the viral proteins. Actin was used as a loading control. VPA‐enhanced virus multiplication in vivo. Viral particles were recovered from aliquots of tumour extracts from animals treated as in panel D, purified by using the QIAamp MinElute Virus Spin Kit (Qiagen) and quantified by real time qPCR. Columns represent the average amounts of the virus recovered from the tumours of three animals with standard deviation bars. Numbers on top of the columns indicate the fold increase in virus titers in the presence versus absence of VPA. 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)