Combination of AT-101/cisplatin overcomes chemoresistance by inducing apoptosis and modulating epigenetics in human ovarian cancer cells
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2013
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Abstract
We investigated the effects of AT-101/cisplatin combination treatment on the expression levels of apoptotic proteins and epigenetic events such as DNA methyltransferase (DNMT) and histone deacetylase (HDAC) enzyme activities in OVCAR-3 and MDAH-2774 ovarian cancer cells. XTT cell viability assay was used to evaluate cytotoxicity. For showing apoptosis, both DNA Fragmentation and caspase 3/7 activity measurements were performed. The expression levels of apoptotic proteins were assessed by human apoptosis antibody array. DNMT and HDAC activities were evaluated by ELISA assay and mRNA levels of DNMT1 and HDAC1 genes were quantified by qRT-PCR. Combination of AT-101/cisplatin resulted in strong synergistic cytotoxicity and apoptosis in human ovarian cancer cells. Combination treatment reduced some pivotal anti-apoptotic proteins such as Bcl-2, HIF-1A, cIAP-1, XIAP in OVCAR-3 cells, whereas p21, Bcl-2, cIAP-1, HSP27, Clusterin and XIAP in MDAH-2774 cells. Among the pro-apoptotic proteins, Bad, Bax, Fas, phospho-p53 (S46), Cleaved caspase-3, SMAC/Diablo, TNFR1 and Cytochrome c were induced in OVCAR-3 cells, whereas, Bax, TRAILR2, FADD, p27, phospho-p53 (S46), Cleaved caspase-3, Cytochrome c, SMAC/Diablo and TNFR1 were induced in MDAH-2774 cells. Combination treatment also inhibited both DNMT and HDAC activities and also mRNA levels in both ovarian cancer cells. AT-101 exhibits great potential in sensitization of human ovarian cancer cells to cisplatin treatment in vitro, suggesting that the combination of AT-101 with cisplatin may hold great promise for development as a novel chemotherapeutic approach to overcome platinum-resistance in human ovarian cancer. © 2012 Springer Science+Business Media Dordrecht.
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Keywords
Antineoplastic Combined Chemotherapy Protocols , Apoptosis , Caspase 3 , Caspase 7 , Cell Line, Tumor , Cell Survival , Cisplatin , DNA (Cytosine-5-)-Methyltransferase , DNA Fragmentation , DNA Methylation , Dose-Response Relationship, Drug , Drug Resistance, Neoplasm , Drug Synergism , Epigenesis, Genetic , Female , Fluorescent Antibody Technique , Gossypol , Histone Deacetylases , Humans , Ovarian Neoplasms , caspase 3 , caspase 7 , cell DNA , cisplatin , clusterin , cytochrome c , death receptor 5 , DNA methyltransferase 1 , Fas antigen , Fas associated death domain protein , heat shock protein 27 , histone deacetylase 1 , hypoxia inducible factor 1alpha , inhibitor of apoptosis protein 1 , isosorbide , messenger RNA , protein BAD , protein Bax , protein bcl 2 , protein p21 , protein p27 , protein p53 , second mitochondrial activator of caspase , tumor necrosis factor receptor 1 , X linked inhibitor of apoptosis , apoptosis , article , cancer cell , cancer resistance , cell strain , cell strain MDAH 2774 , cell strain OVCAR 3 , cell viability , chemosensitization , controlled study , DNA fragmentation , DNMT1 gene , drug cytotoxicity , drug potentiation , enzyme activity , enzyme inhibition , epigenetics , HDAC1 gene , human , human cell , in vitro study , ovary cancer , protein cleavage , protein expression