Simultaneous modulation of the intrinsic and extrinsic pathways by simvastatin in mediating prostate cancer cell apoptosis

BMC Cancer

Anna Goc³  Samith T Kochuparambil²  Belal Al-Husein³  Ahmad Al-Azayzih³  Shuaib Mohammad⁴  Payaningal R Somanath¹ 

[1] Clinical and Experimental Therapeutics, College of Pharmacy, University of Georgia, HM1200 – Georgia Health Sciences University, Augusta, GA, 30912, USA

[2] Department of Medicine, Georgia Health Sciences University, Augusta, GA, USA

[3] Charlie Norwood VA Medical Center, Augusta, GA, USA

[4] Clinical and Experimental Therapeutics, College of Pharmacy, University of Georgia, Augusta, GA, USA

关键词: Fas-L;    Bcl-2;    Apoptosis;    Docetaxel;    Simvastatin;    Prostate cancer;   

Others  :  1080200
DOI  :  10.1186/1471-2407-12-409

 received in 2012-04-14, accepted in 2012-09-11,  发布年份 2012

【 摘 要 】

Background

Recent studies suggest the potential benefits of statins as anti-cancer agents. Mechanisms by which statins induce apoptosis in cancer cells are not clear. We previously showed that simvastatin inhibit prostate cancer cell functions and tumor growth. Molecular mechanisms by which simvastatin induce apoptosis in prostate cancer cells is not completely understood.

Methods

Effect of simvastatin on PC3 cell apoptosis was compared with docetaxel using apoptosis, TUNEL and trypan blue viability assays. Protein expression of major candidates of the intrinsic pathway downstream of simvastatin-mediated Akt inactivation was analyzed. Gene arrays and western analysis of PC3 cells and tumor lysates were performed to identify the candidate genes mediating extrinsic apoptosis pathway by simvastatin.

Results

Data indicated that simvastatin inhibited intrinsic cell survival pathway in PC3 cells by enhancing phosphorylation of Bad, reducing the protein expression of Bcl-2, Bcl-xL and cleaved caspases 9/3. Over-expression of PC3 cells with Bcl-2 or DN-caspase 9 did not rescue the simvastatin-induced apoptosis. Simvastatin treatment resulted in increased mRNA and protein expression of molecules such as TNF, Fas-L, Traf1 and cleaved caspase 8, major mediators of intrinsic apoptosis pathway and reduced protein levels of pro-survival genes Lhx4 and Nme5.

Conclusions

Our study provides the first report that simvastatin simultaneously modulates intrinsic and extrinsic pathways in the regulation of prostate cancer cell apoptosis in vitro and in vivo, and render reasonable optimism that statins could become an attractive anti-cancer agent.

【 授权许可】

   
2012 Goc et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

Figure 9.

【 参考文献 】

• [1]Jakobisiak M, Golab J: Statins can modulate effectiveness of antitumor therapeutic modalities. Med Res Rev 2010, 30:102-135.
• [2]Liao JK, Laufs U: Pleiotropic effects of statins. Annu Rev Pharmacol Toxicol 2005, 45:89-118.
• [3]Hamilton RJ, Banez LL, Aronson WJ, Terris MK, Platz EA, Kane CJ, Presti JC Jr, Amling CL, Freedland SJ: Statin medication use and the risk of biochemical recurrence after radical prostatectomy: results from the Shared Equal Access Regional Cancer Hospital (SEARCH) Database. Cancer 2010, 116:3389-3398.
• [4]Marcella SW, David A, Ohman-Strickland PA, Carson J, Rhoads GG: Statin use and fatal prostate cancer: A matched case–control study. Cancer 2012, 118(16):4046-4052.
• [5]Kochuparambil ST, Al-Husein B, Goc A, Soliman S, Somanath PR: Anticancer efficacy of simvastatin on prostate cancer cells and tumor xenografts is associated with inhibition of Akt and reduced prostate-specific antigen expression. J Pharmacol Exp Ther 2011, 336:496-505.
• [6]Spampanato C, De Maria S, Sarnataro M, Giordano E, Zanfardino M, Baiano S, Carteni M, Morelli F: Simvastatin inhibits cancer cell growth by inducing apoptosis correlated expression. Int J Oncol 2012, 40:935-941.
• [7]Fulda S, Debatin KM: 5-Aza-2′-deoxycytidine and IFN-gamma cooperate to sensitize for TRAIL-induced apoptosis by upregulating caspase-8. Oncogene 2006, 25:5125-5133.
• [8]French LE, Tschopp J: Protein-based therapeutic approaches targeting death receptors. Cell Death Differ 2003, 10:117-123.
• [9]Wajant H: Death receptors. Essays Biochem 2003, 39:53-71.
• [10]Peter ME, Krammer PH: The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ 2003, 10:26-35.
• [11]Micheau O, Tschopp J: Induction of TNF receptor I-mediated apoptosis via two sequential signaling complexes. Cell 2003, 114:181-190.
• [12]Massard C, Fizazi K: Targeting continued androgen receptor signaling in prostate cancer. Clin Cancer Res 2011, 17:3876-3883.
• [13]Huggins C, Hodges CV: Studies on prostatic cancer: I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. 1941. J Urol 2002, 168:9-12.
• [14]Herbst RS, Khuri FR: Mode of action of docetaxel - a basis for combination with novel anticancer agents. Cancer Treat Rev 2003, 29:407-415.
• [15]Eckhoff L, Nielsen M, Moeller S, Knoop A: TAXTOX - a retrospective study regarding the side effects of docetaxel given as part of the adjuvant treatment to patients with primary breast cancer in Denmark from 2007 to 2009. Acta Oncol 2011, 50:1075-1082.
• [16]Pivot X, Schneeweiss A, Verma S, Thomssen C, Passos-Coelho JL, Benedetti G, Ciruelos E, von Moos R, Chang HT, Duenne AA, Miles DW: Efficacy and safety of bevacizumab in combination with docetaxel for the first-line treatment of elderly patients with locally recurrent or metastatic breast cancer: results from AVADO. Eur J Cancer 2011, 47:2387-2395.
• [17]Thibault A, Samid D, Tompkins AC, Figg WD, Cooper MR, Hohl RJ, Trepel J, Liang B, Patronas N, Venzon DJ, et al.: Phase I study of lovastatin, an inhibitor of the mevalonate pathway, in patients with cancer. Clin Cancer Res 1996, 2:483-491.
• [18]van der Spek E, Bloem AC, van de Donk NW, Bogers LH, van der Griend R, Kramer MH, de Weerdt O, Wittebol S, Lokhorst HM: Dose-finding study of high-dose simvastatin combined with standard chemotherapy in patients with relapsed or refractory myeloma or lymphoma. Haematologica 2006, 91:542-545.
• [19]Holstein SA, Knapp HR, Clamon GH, Murry DJ, Hohl RJ: Pharmacodynamic effects of high dose lovastatin in subjects with advanced malignancies. Cancer Chemother Pharmacol 2006, 57:155-164.
• [20]Somanath PR, Vijai J, Kichina JV, Byzova T, Kandel ES: The role of PAK-1 in activation of MAP kinase cascade and oncogenic transformation by Akt. Oncogene 2009, 28:2365-2369.
• [21]Goc A, Al-Husein B, Kochuparambil ST, Liu J, Heston WW, Somanath PR: PI3 kinase integrates Akt and MAP kinase signaling pathways in the regulation of prostate cancer. Int J Oncol 2011, 38:267-277.
• [22]Crowell JA, Steele VE, Fay JR: Targeting the AKT protein kinase for cancer chemoprevention. Mol Cancer Ther 2007, 6:2139-2148.
• [23]Davies MA: Regulation, role, and targeting of Akt in cancer. J Clin Oncol 2011, 29:4715-4717.
• [24]Chen J, Somanath PR, Razorenova O, Chen WS, Hay N, Bornstein P, Byzova TV: Akt1 regulates pathological angiogenesis, vascular maturation and permeability in vivo. Nat Med 2005, 11:1188-1196.
• [25]Somanath PR, Kandel ES, Hay N, Byzova TV: Akt1 signaling regulates integrin activation, matrix recognition, and fibronectin assembly. J Biol Chem 2007, 282:22964-22976.
• [26]Somanath PR, Razorenova OV, Chen J, Byzova TV: Akt1 in endothelial cell and angiogenesis. Cell Cycle 2006, 5:512-518.
• [27]Shibata MA, Ito Y, Morimoto J, Otsuki Y: Lovastatin inhibits tumor growth and lung metastasis in mouse mammary carcinoma model: a p53-independent mitochondrial-mediated apoptotic mechanism. Carcinogenesis 2004, 25:1887-1898.
• [28]Gauthaman K, Manasi N, Bongso A: Statins inhibit the growth of variant human embryonic stem cells and cancer cells in vitro but not normal human embryonic stem cells. Br J Pharmacol 2009, 157:962-973.
• [29]Fukuda S, Pelus LM: Survivin, a cancer target with an emerging role in normal adult tissues. Mol Cancer Ther 2006, 5:1087-1098.
• [30]Fornaro M, Plescia J, Chheang S, Tallini G, Zhu YM, King M, Altieri DC, Languino LR: Fibronectin protects prostate cancer cells from tumor necrosis factor-alpha-induced apoptosis via the AKT/survivin pathway. J Biol Chem 2003, 278:50402-50411.
• [31]Krajewska M, Krajewski S, Banares S, Huang X, Turner B, Bubendorf L, Kallioniemi OP, Shabaik A, Vitiello A, Peehl D, et al.: Elevated expression of inhibitor of apoptosis proteins in prostate cancer. Clin Cancer Res 2003, 9:4914-4925.
• [32]Amaravadi R, Thompson CB: The survival kinases Akt and Pim as potential pharmacological targets. J Clin Invest 2005, 115:2618-2624.
• [33]Vaira V, Lee CW, Goel HL, Bosari S, Languino LR, Altieri DC: Regulation of survivin expression by IGF-1/mTOR signaling. Oncogene 2007, 26:2678-2684.
• [34]Munier A, Feral C, Milon L, Pinon VP, Gyapay G, Capeau J, Guellaen G, Lacombe ML: A new human nm23 homologue (nm23-H5) specifically expressed in testis germinal cells. FEBS Lett 1998, 434:289-294.
• [35]Kawamata N, Sakajiri S, Sugimoto KJ, Isobe Y, Kobayashi H, Oshimi K: A novel chromosomal translocation t(1;14)(q25;q32) in pre-B acute lymphoblastic leukemia involves the LIM homeodomain protein gene, Lhx4. Oncogene 2002, 21:4983-4991.
• [36]Hung TM, Hu RH, Ho CM, Chiu YL, Lee JL, Jeng YM, Shih DT, Lee PH: Downregulation of alpha-fetoprotein expression by LHX4: a critical role in hepatocarcinogenesis. Carcinogenesis 2011, 32:1815-1823.
• [37]Bhardwaj A, Aggarwal BB: Receptor-mediated choreography of life and death. J Clin Immunol 2003, 23:317-332.
• [38]Sprick MR, Rieser E, Stahl H, Grosse-Wilde A, Weigand MA, Walczak H: Caspase-10 is recruited to and activated at the native TRAIL and CD95 death-inducing signalling complexes in a FADD-dependent manner but can not functionally substitute caspase-8. EMBO J 2002, 21:4520-4530.
• [39]Varfolomeev EE, Ashkenazi A: Tumor necrosis factor: an apoptosis JuNKie? Cell 2004, 116:491-497.