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Targeting Redox Homeostasis in Malignant Mesothelioma

Kheng Newick1, Brian Cunniff1, Paul Held2, Arti Shukla1, Harvey Pass3, Kelly Butnor1, Kousik Kundu4, Niren Murthy4, Brooke Mossman1, Jack Arbiser5, and Nicholas Heintz1 1 2 3 University of Vermont, Biotek Instruments, New York 4 5 University, Georgia Institute of Technology, Emory University Effective therapy is lacking for malignant mesothelioma (MM), a highly aggressive cancer linked to occupational asbestos exposure. Redox-dependent signaling by reactive oxygen species (ROS) plays an important role in cancer pathogenesis, and may represent a therapeutic target in MM. We have explored the role of FOXM1, a redox-dependent transcription factor that regulates resistance to oxidative stress and entry into G2/M in MM cell proliferation and viability. FOXM1 is expressed at higher levels in MM cells and, in contrast to control telomeraseimmortalized mesothelial cell line LP9, continues to be expressed in the absence of growth factors. Human MM tumors express more FOXM1 transcript than normal mesothelial tissue, and immunostaining of human MM tissue arrays confirms that FOXM1 is highly expressed in human MM. Studies in vitro show that MM cells constitutively generate approximately twice the level of superoxide, which is predominantly derived from mitochondria. the triphenylmethane gentian violet (GV) and the thiazole antibiotic thiostrepton (TS) inhibit FOXM1 expression, MM tumor cell viability and impede tumor progression in mouse xenograft models of human MM. While GV induces mitochondrial depolarization and inhibits the production of mitochondrial ROS, and TS hyperpolarizes the mitochondria and increases mitochondrial ROS production, both compounds extinguished the expression of FOXM1 mRNA and protein in a dose-dependent manner. TS, but not GV, alters the electrophoretic mobility of the mitochondrial antioxidant enzyme peroxiredoxin 3 (PRX3), possibly by adducting the peroxidatic cysteine, thereby inducing intolerable levels of mitochondrial oxidative stress. Interestingly GV markedly potentiates the effect of TS on PRX3. Taken together, our studies indicate that expression of FOXM1 is tuned to mitochondrial oxidant production, and compounds that target the redox homeostasis of mitochondria may be useful in the treatment of MM.

together, our results suggest that the suppression of IDPm activity resulted in the disruption of cellular redox balance and subsequently exacerbates EGCG-induced apoptotic cell death in HeLa cells. These results might have implications for developing an effective combination modality in cancer treatment.

doi:10.1016/j.freeradbiomed.2011.10.333

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Oxidative Stress Induction and Antitumor Effect of Two Fenilaminonaftoquinones Against the Ehrlich Ascites Carcinoma
Karina Bettega Felipe1, Mirelle S Farias1, Tania M F Gunther1, Nadia F Bucker1, Maicon Roberto Kviecinski1, Fabiana Ourique1, Valdelucia V.M.A.S Grinevicius1, Geremias Reginaldo1, Benites J2, Pedro Buc Calderon3, and Rozangela Curi Pedrosa1 1 2 universidade Federal De Santa Catarina, Departamento de Ciencias Qumicas y Farmacuticas, Universidad Arturo Prat, 3 Iquique, Toxicology and Cancer Biology Research Group, Louvain Drug Research, Institute Universit Catholique Objective: Synthetic quinones have been researched for cancer chemotherapy, since some of them have potential antitumor activities, as well doxorubicin belong to the same drug class and a chemotherapeutic agent, whose action is also due to its ability to generate free radicals. the aim of this work was to verify if the antitumor effect of two synthetic quinones (Q7 and Q7A) could be related to oxidative stress induction. Methods: EhrIich ascites carcinoma (EAC) was inoculated in isogenic Balb/c male mice (~20g b.w., n=6) and this moment was taken as day 0. Intraperitoneal treatments with Q7 and Q7A (1mg/Kg per day) started on the first day and on tenth the animals were sacrificed. Negative Control (NC) received only saline. Their ascitic fluid was collected and used for the analysis. the following parameters were assessed: inhibition of tumor growth, percent of necrotic and apoptotic cells, as well as lipid peroxidation, activities of superoxide dismutase and catalase. Results were expressed by means and standard deviation and were analyzed using one-way ANOVA and Tukey-Kramers test. a P-value < 0.05 was considered to be statistically significant. Results: Q7 and Q7A decreased the tumor growth(Q7= 64.3; Q7A= 57.2; NC=0%), and increased the percent of necrotic cells(Q7= 19.5; Q7A= 45.5; NC=2.4%) when compared to NC. Only Q7 was able to increase the levels of lipid peroxidation(Q7= 14.3 3.6; Q7A= 8.5 1.4; NC= 6.2 4,0 mol/mg protein). Furthermore, the quinone treatments increased the superoxide dismutase(Q7= 71.3 4.6; Q7A= 27.8 2.0; NC= 13.2 5.0 USOD/mg protein) and catalase activities (Q7= 9.33.4; Q7A= 27.91.2; NC= 2.92.3 min/ mg protein). Conclusion: These findings suggest that the antitumor activity of Q7 and Q7A could be related to induction of free radical generation. Keywords: Fenilaminonaftoquinones; antitumor effect; ehrlich ascites carcinoma

doi:10.1016/j.freeradbiomed.2011.10.332

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Attenuated mitochondrial NADP+-dependent isocitrate dehydrogenase activity enhances EGCGinduced apoptosis
Kyu Ho Jung1, Kil in Sup1, and Jeen-Woo Park1 1 Kyungpook National University (-)-Epigallocatechin-3-gallate (EGCG), a well-known chemopreventive factor, induces cancer cells undergoing apoptosis. Over the last several years, we have shown that the + mitochondrial NADP -dependent isocitrate dehydrogenase (IDPm) functions as an antioxidant and anti-apoptotic protein by supplying NADPH to antioxidant systems. Here, we show that EGCG induced the inactivation of IDPm as a purified enzyme and in cultured cancer cells in a dose- and time-dependent manner. Loss of enzyme activity was associated with the depletion of the thiol groups in protein. in addition, transfection of HeLa cells with an IDPm small interfering RNA (siRNA) markedly attenuated the activity of IDPm and substantially enhanced EGCG-induced apoptosis as indicated by the morphological evidence of apoptosis, DNA fragmentation, and the modulation of mitochondrial function and apoptotic marker proteins. Taken

doi:10.1016/j.freeradbiomed.2011.10.334

SFRBM 2011

S127