Proteasome and HDAC Inhibition Changes the Expression Levels of Bcl-2, Bcl-XL, Bim and Bik Proteins in Androgen-Independent PC-3 Cell Line
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.
References
Shen MM, Abate-Shen C. Molecular genetics of prostate cancer: new prospects for old challenges. Genes Dev. 2010; 24: 1967-2000.
Rider JR, Sandin F, Andrén O, Wiklund P, Hugosson J, Stattin P. Long-term outcomes among noncuratively treated men according to prostate cancer risk category in a nationwide, population-based study. Eur Urol. 2013; 63:88-96.
Chen D, Frezza M, Schmitt S, Kanwar J, Dou QP. Bortezomib as the first proteasome inhibitor anticancer drug: current status and future perspectives. Curr Cancer Drug Targets. 2011; 11: 239-53.
Frankland-Searby S, Bhaumik SR. The 26S proteasome complex: an attractive target for cancer therapy. Biochim Biophys Acta. 2012; 1825: 64-76.
Carew JS, Giles FJ, Nawrocki ST. Histone deacetylase inhibitors: mechanisms of cell death and promise in combination cancer therapy. Cancer Letters. 2008; 269: 7-17.
Lin Z, Bazzaro M, Wang MC, Chan KC, Peng S, Roden R. Combination of proteasome and HDAC inhibitors for uterine cervical cancer treatment. Clin Cancer Res. 2009; 15: 570–7.
Heider U, Rademacher J, Lamottke B, Mieth M, Moebs M, von Metzler I, et al. Synergistic interaction of the histone deacetylase inhibitor SAHA with the proteasome inhibitor bortezomib in cutaneous T cell lymphoma. Eur J Haematol. 2009; 82: 440–9.
Ellis L, Pili R. Histone deacetylase inhibitors: advancing therapeutic strategies in hematological and solid malignancies. Pharmaceuticals 2010; 3(8): 2411-69.
Brunelle JK, Leatai A. Control of mitochondrial apoptosis by the Bcl-2 family. J Cell Sci. 2009; 122 (Pt 4): 437-41.
Fennell DA, Chacko A, Mutti L. Bcl-2 family regulation by the 20S proteasome inhibitor bortezomib. Oncogene 2008; 27: 1189–97.
Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol. 2008; 9:47-59.
Brancolini C. Inhibitors of the ubiquitin-proteasome system and the cell death machinery: how many pathways are activated? Curr Mol Pharmacol. 2008; 1: 24-37.
Yip KW, Reed JC. Bcl-2 family proteins and cancer. Oncogene 2008; 27: 6398-406.
Gurumurthy S, Vasudevan KM, Rangnekar VM. Regulation of apoptosis in prostate cancer. Cancer Metastasis Rev. 2001; 20: 225-43.
Vela L, Marzo I. Bcl-2 family of proteins as drug targets for cancer chemotherapy: the long way of BH3 mimetics from bench to bedside. Curr Opin Pharmacol. 2015; 23:74-81.
McConkey DJ, Zhu K. Mechanisms of proteasome inhibitor action and resistance in cancer. Drug Resist Updat. 2008; 11: 164-79.
Marshansky V, Wang X, Bertrand R, Luo H, Duguid W, Chinnadurai G, et al. Proteasomes modulate balance among proapoptotic and antiapoptotic Bcl-2 family members and compromise functioning of the electron transportchain in leukemic cells. J Immunol. 2001; 166: 3130-42.
Nikrad M, Johnson T, Puthalalath H, Coultas L, Adams J, Kraft AS. The proteasome inhibitor bortezomib sensitizes cells to killing by death receptor ligand TRAIL via BH3-only proteins Bik and Bim. Mol Cancer Ther. 2005;4:443-9.
Fang Y, Hu Y, Wu P, Wang B, Tian Y, Xia X, et al. Synergistic efficacy in human ovarian cancer cells by histone deacetylase inhibitor TSA and proteasome inhibitor PS-341. Cancer Invest. 2011; 29: 247-52.
Pei XY, Dai Y, Grant S. Synergistic induction of oxidative injury and apoptosis in human multiple myeloma cells by the proteasome inhibitor bortezomib and histone deacetylase inhibitors. Clinical Cancer Research 2004; 10: 3839-52.
Denlinger CE, Keller MD, Mayo MW, Broad RM, Jones DR. Combined proteasome and histone deacetylase inhibition in non-small cell lung cancer. J Thorac Cardiovasc Surg. 2004; 127: 1078-86.
Kim J, Guan J, Chang I, Chen X, Han D, Wang CY. PS-341 and histone deacetylase inhibitor synergistically induce apoptosis in head and neck squamous cell carcinoma cells. Mol Cancer Ther. 2010; 9: 1977-84.
