Abstract
This study evaluates the effect of simultaneous AKT inhibition and cisplatin therapy in changes of Reactive Oxygen Species (ROS) production, apoptosis induction, and cell survival in cisplatin-resistant OVCAR3 cell. OVCAR3 cancer cells were treated with cisplatin, Ly 294002 (LY), and cisplatin+Ly to investigate the cytotoxicity effect of the mentioned groups via MTT assay. Then, DCFH-DA (2', 7'-dichlorodihydro fluorescein diacetate) assay kit is used to assess the potential of treated groups in intracellular ROS generation. Protein expression levels of caspase-3, cleaved caspase 3, PI3K, Akt, p-Akt, XIAP, and Survivin are estimated through immunoblotting assay in all three experimental groups. The results showed that all three treated groups, including cisplatin and Ly alone and co-administration of cisplatin+Ly, could reduce the cell vitality of OVCAR3 cancer cells, induced intracellular production of ROS and increased the expression level of activated caspase 3 and Akt protein, whereas down-regulated the phosphorylation of Akt protein. However, the effect of combination therapy was more tangible compared to single therapy and control groups. In contrast, the expression amount of XIAP, Survivin, and PI3K did not show detectable changes in comparison with the control group. The results showed that the AKT inhibition by Ly could sensitize the OVCAR3 cancer cells to the cisplatin and lower the effective dose of cisplatin through hyperactivation of oxidative stress.
Highlights
Ovarian cancer with the fourth most common cause of cancer-related death can be considered as an aggressive and under-recognized gynecological malignancy in women across the world [1-3]
We aimed to investigate the effects of Ly and cisplatin either alone or simultaneously on Reactive Oxygen Species (ROS) generation, apoptosis-related proteins, caspase 3 and xXIAP activation, and cell survival-related proteins Phosphatidylinositol 3-kinase (PI3K), AKT and surviving regulation
Cis+Ly reduced cell viability more than single therapy with an IC50 amount of 5.46 μM, 4.60 μM, and 4.82 μM during 24, 48, and 72 h, respectively, which proved the increased sensitivity of OVCAR3 cells to cisplatin in the presence of 5 uM of Ly (Table 1)
Summary
Ovarian cancer with the fourth most common cause of cancer-related death can be considered as an aggressive and under-recognized gynecological malignancy in women across the world [1-3]. This study evaluates the effect of simultaneous AKT inhibition and cisplatin therapy in changes of Reactive Oxygen Species (ROS) production, apoptosis induction, and cell survival in cisplatinresistant OVCAR3 cell. Methods: OVCAR3 cancer cells were treated with cisplatin, Ly 294002 (LY), and cisplatin+Ly to investigate the cytotoxicity effect of the mentioned groups via MTT assay. Results: The results showed that all three treated groups, including cisplatin and Ly alone and coadministration of cisplatin+Ly, could reduce the cell vitality of OVCAR3 cancer cells, induced intracellular production of ROS and increased the expression level of activated caspase 3 and Akt protein, whereas downregulated the phosphorylation of Akt protein. Conclusions: The results showed that the AKT inhibition by Ly could sensitize the OVCAR3 cancer cells to the cisplatin and lower the effective dose of cisplatin through hyperactivation of oxidative stress
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