%0 Journal Article
%T Sensitisation to mitoxantrone-induced apoptosis by the oncolytic adenovirus Ad£¿£¿ through Bcl-2-dependent attenuation of autophagy
%A Carmen Aguirre-Hern¨¢ndez
%A Gunnel Halld¨¦n
%A H¨¦ctor Maya-Pineda
%A Julia San Mill¨¢n
%A Y. K. Stella Man
%A Yong-Jie Lu
%J Archive of "Oncogenesis".
%D 2018
%R 10.1038/s41389-017-0020-8
%X a Sensitisation to mitoxantrone by Ad5wt, Ad£¿£¿ and AdE1A12S in PC3 and 22Rv1 cells. Dose£¿response curves to mitoxantrone with and without fixed doses of virus killing 10¨C40% of cells alone; Ad5wt or Ad¦¤¦¤ (1000£¿ppc; PC3 and 25£¿ppc;22Rv1) or AdE1A12S (5000£¿ppc;PC3and 100£¿ppc;22Rv1). Cell viability determined by MTS assays 5 (PC3) or 3 days (22Rv1) after treatment and data analysed by unpaired t-test, averages£¿¡À£¿SEM, n£¿¡Ý£¿3, **p£¿<£¿0.01, *p£¿<£¿0.05. Ratios£¿=£¿EC50-values of combination/EC50-values mitoxantrone. b PC3 and 22Rv1 cells infected with Ad£¿£¿ or AdE1A12S and treated with mitoxantrone for 5 and 3 days, respectively, at three constant ratios (0.5, 2.5 and 12.5 ppc/nm; indicated by wedges) and PC3M cells at two constant ratios (2.5 and 12.5£¿ppc/nm). Combination indexes (CI) calculated from isobolograms and synergistic cell killing defined as CI£¿¡Ü£¿0.9; averages£¿¡À£¿SD, n£¿=£¿3, *p£¿<£¿0.05 compared to the theoretical additive values (dashed lines; additive effects 0.9£¿<£¿CI£¿<£¿1.1). c PC3 cells (left panel) treated with mitoxantrone (225, 450 and 900£¿nm) and/or infected with Adwt and Ad¦¤¦¤ at 500ppc or Ad12S at 5000ppc. Cell death indirectly determined using the MTS viability assay 5 days after treatment. One-way ANOVA with Tukey£¿Kramer post-test, averages£¿¡À£¿SEM, n£¿=£¿4. 22Rv1 cells (right panel) treated with mitoxantrone (3, 10 and 25£¿nm) and/or infected with Ad5wt and Ad¦¤¦¤ at 15£¿ppc or Ad12S at 50£¿ppc. Number of dead cells determined by the Trypan blue exclusion assay after 3 days. Representative study, one-way ANOVA with Tukey£¿Kramer post-test, averages£¿¡À£¿SEM from quadruplicate samples. *p£¿<£¿0.05, ***p£¿<£¿0.001, ****p£¿<£¿0.0001, *compared to the theoretical additive value of mitoxantrone and virus, n£¿¡Ý£¿3. d Expression of PARP detected by immunoblotting (cleaved PARP indicated with black arrows; 89£¿kDa), one representative immunoblot, n£¿=£¿3. PC3 cells (left panel) infected with Ad¦¤¦¤ (500£¿ppc) or AdE1A12S (5000£¿ppc) and/or mitoxantrone (225, 450 or 900£¿nm). 22Rv1 cells (right panel) infected with Ad¦¤¦¤ (15£¿ppc) or AdE1A12S (50£¿ppc) and/or mitoxantrone (3, 10 or 25£¿nm). Cells were lysed 48£¿h after treatment. Lower panels: PARP ratios; cleaved PARP (clPARP/PARP) after quantification by densitometry and normalised to the actin loading control, expressed as fold-change relative to the untreated control, n£¿=£¿3. Molecular weight markers indicated in kDa. e PC3 cells analysed at the indicated time points for mitochondrial depolarisation by loss of TMRE staining using flow cytometry (left panel) and Annexin V staining (right panel) 120£¿h after infection with Ad¦¤¦¤ (500£¿ppc)
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833340/