Abstract
Objective: Prostate cancer is the most common cancer observed and it is estimated to have caused 10% of all cancer-related deaths in men in western countries. Transition from androgen-dependent form to metastatic androgen-independent phase is a consequence of deranged apoptotic response by the cancer cells to therapy. Bcl-2 family members are important modulators of apoptotic response. In this study, we investigated the effects of proteasome and histone deacetylase (HDAC) inhibitor treatment on the expression changes in some Bcl-2 family genes at protein level. Methods: PC-3 cells were cultured in plate before being exposed to different concentrations of unaccompanied bortezomib and TSA as well as bortezomib – TSA combination. After the protein isolation from control and treated cells, whole cell lysate was used for determining Bcl-2, Bcl-XL, Bim and Bik proteins with western blotting method. Results: We observed that after the drug treatment, the expression levels of pro-apoptotic Bim and Bik proteins have increased and those of anti-apoptotic Bcl-2 and Bcl-XL proteins have decreased. Synergy between bortezomib and TSA induces apoptosis. Conclusion: Our results showed that application of low doses of bortezomib and TSA combination may be sufficient for apoptotic response. Our findings may come in handy for the advanced prostate cancer research.
Highlights
MethodsCell Culture and Reagents The human prostate cancer cell line PC-3 was provided by Prof
We examined the anti-apoptotic effects of unaccompanied bortezomib and TSA as well as bortezomib – TSA combination on the human PC3 prostate cancer cell line and investigated the expression levels of antiapoptotic Bcl-2, Bcl-XL and pro-apoptotic Bim and Bik proteins
To identify the changes in the expression levels of anti-apoptotic Bcl-2, BclXL and pro-apoptotic Bim and Bik, protein lysates were obtained from PC-3 cells which underwent unaccompanied bortezomib (10nM and 50nM) and TSA (400nM) as well as combination (5nM Bortezomib and 300nM TSA) treatment
Summary
Cell Culture and Reagents The human prostate cancer cell line PC-3 was provided by Prof. Levent Türkeri (Department of Urology, Faculty of Medicine, Marmara University, Istanbul, Turkey). Cells were cultured in RPMI medium containing 10% fetal bovine serum and 1% penicillin-streptomycin and maintained at 37°C with 5% carbon dioxide. Bortezomib was purchased from BioVision (Milpitas, CA, USA). It was dissolved in dimethylsulfoxide at a concentration of 1mM and stored at -20°C. TSA was purchased from Cell Signaling Technology Inc. It was dissolved in ethanol at a 1mM stock solution and stored at -20°C. Stock solutions were diluted to their working concentrations with growth medium just before use
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