Combined pitavastatin and dacarbazine treatment activates apoptosis and autophagy resulting in synergistic cytotoxicity in melanoma cells

Al-Qatati,Abeer; Aliwaini,Saeb

doi:10.3892/ol.2017.7189

Combined pitavastatin and dacarbazine treatment activates apoptosis and autophagy resulting in synergistic cytotoxicity in melanoma cells

Authors:
- Abeer Al-Qatati
- Saeb Aliwaini
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Affiliations: Faculty of Applied Medical Sciences, Al-Azhar University, Gaza 1277, Palestine, Department of Biological Sciences and Biotechnology, Faculty of Sciences, Islamic University of Gaza, Gaza 108, Palestine
Published online on: October 16, 2017 https://doi.org/10.3892/ol.2017.7189
Pages: 7993-7999

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Abstract

Melanoma is an aggressive skin cancer and its incidence is increasing faster than any other type of cancer. Whilst dacarbazine (DTIC) is the standard chemotherapy for metastatic melanoma, it has limited success. Statins, including pitavastatin, have been demonstrated to have a range of anti-cancer effects in a number of human cancer cell lines. The present study therefore explored the anti-cancer activity of combined DTIC and pitavastatin in A375 and WM115 human melanoma cells. Cell survival assays demonstrated that combined DTIC and pitavastatin treatment resulted in synergistic cell death. Cell cycle analyses further revealed that this combined treatment resulted in a G1 cell cycle arrest, as well as a sub-G1 population, indicative of apoptosis. Activation of apoptosis was confirmed by Annexin V-fluorescein isothiocyanate/propidium iodide double-staining and an increase in the levels of active caspase 3 and cleaved poly (ADP-ribose) polymerase. Furthermore, it was demonstrated that apoptosis occurs through the intrinsic pathway, evident from the release of cytochrome c. Finally, combined DTIC and pitavastatin treatment was demonstrated to also activate autophagy as part of a cell death mechanism. The present study provides novel evidence to suggest that the combined treatment of DTIC and pitavastatin may be effective in the treatment of melanoma.

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1	Bhatia S, Tykodi SS and Thompson JA: Treatment of metastatic melanoma: An overview. Oncology (Williston Park). 23:488–496. 2009.PubMed/NCBI
2	Li W and Melton DW: Cisplatin regulates the MAPK kinase pathway to induce increased expression of DNA repair gene ERCC1 and increase melanoma chemoresistance. Oncogene. 31:2412–2422. 2012. View Article : Google Scholar : PubMed/NCBI
3	Lui P, Cashin R, Machado M, Hemels M, Corey-Lisle PK and Einarson TR: Treatments for metastatic melanoma: Synthesis of evidence from randomized trials. Cancer Treat Rev. 33:665–680. 2007. View Article : Google Scholar : PubMed/NCBI
4	Bedikian AY, Millward M, Pehamberger H, Conry R, Gore M, Trefzer U, Pavlick AC, DeConti R, Hersh EM, Hersey P, et al: Bcl-2 antisense (oblimersen sodium) plus dacarbazine in patients with advanced melanoma: The Oblimersen Melanoma Study Group. J Clin Oncol. 24:4738–4745. 2006. View Article : Google Scholar : PubMed/NCBI
5	Lev DC, Onn A, Melinkova VO, Miller C, Stone V, Ruiz M, McGary EC, Ananthaswamy HN, Price JE and Bar-Eli M: Exposure of melanoma cells to dacarbazine results in enhanced tumor growth and metastasis in vivo. J Clin Oncol. 22:2092–2100. 2004. View Article : Google Scholar : PubMed/NCBI
6	Gopalan A, Yu W, Sanders BG and Kline K: Simvastatin inhibition of mevalonate pathway induces apoptosis in human breast cancer cells via activation of JNK/CHOP/DR5 signaling pathway. Cancer Lett. 329:9–16. 2013. View Article : Google Scholar : PubMed/NCBI
7	Warita K, Warita T, Beckwitt CH, Schurdak ME, Vazquez A, Wells A and Oltvai ZN: Statin-induced mevalonate pathway inhibition attenuates the growth of mesenchymal-like cancer cells that lack functional E-cadherin mediated cell cohesion. Sci Rep. 4:75932014. View Article : Google Scholar : PubMed/NCBI
8	Simon A: Cholesterol metabolism and immunity. N Engl J Med. 371:1933–1935. 2014. View Article : Google Scholar : PubMed/NCBI
9	Tu YS, Kang XL, Zhou JG, Lv XF, Tang YB and Guan YY: Involvement of Chk1-Cdc25A-cyclin A/CDk2 pathway in simvastatin induced S-phase cell cycle arrest and apoptosis in multiple myeloma cells. Eur J Pharmacol. 670:356–364. 2011. View Article : Google Scholar : PubMed/NCBI
10	Lee YP, Wang CW, Liao WC, Yang CR, Yeh CT, Tsai CH, Yang CC and Tzeng YM: In vitro and in vivo anticancer effects of mevalonate pathway modulation on human cancer cells. Anticancer Res. 32:2735–2745. 2012.PubMed/NCBI
11	McFarland AJ, Anoopkumar-Dukie S, Arora DS, Grant GD, McDermott CM, Perkins AV and Davey AK: Molecular mechanisms underlying the effects of statins in the central nervous system. Int J Mol Sci. 15:20607–20637. 2014. View Article : Google Scholar : PubMed/NCBI
12	Garwood ER, Kumar AS, Baehner FL, Moore DH, Au A, Hylton N, Flowers CI, Garber J, Lesnikoski BA, Hwang ES, et al: Fluvastatin reduces proliferation and increases apoptosis in women with high grade breast cancer. Breast Cancer Res Treat. 119:137–144. 2010. View Article : Google Scholar : PubMed/NCBI
13	Wang J, Tokoro T, Higa S and Kitajima I: Anti-inflammatory effect of pitavastatin on NF-kappaB activated by TNF-alpha in hepatocellular carcinoma cells. Biol Pharm Bull. 29:634–639. 2006. View Article : Google Scholar : PubMed/NCBI
14	Fujino M, Miura S, Matsuo Y, Tanigawa H, Kawamura A and Saku K: Pitavastatin-induced downregulation of CCR2 and CCR5 in monocytes is associated with the arrest of cell-cycle in S phase. Atherosclerosis. 187:301–308. 2006. View Article : Google Scholar : PubMed/NCBI
15	Tsuboi Y, Kurimoto M, Nagai S, Hayakawa Y, Kamiyama H, Hayashi N, Kitajima I and Endo S: Induction of autophagic cell death and radiosensitization by the pharmacological inhibition of nuclear factor-kappa B activation in human glioma cell lines. J Neuosurg. 110:594–604. 2009. View Article : Google Scholar
16	Aliwaini S, Swarts AJ, Blanckenberg A, Mapolie S and Prince S: A novel binuclear palladacycle complex inhibits melanoma growth in vitro and in vivo through apoptosis and autophagy. Biochem Pharmacol. 86:1650–1663. 2013. View Article : Google Scholar : PubMed/NCBI
17	Bagnoli M, Balladore E, Luison E, Alberti P, Raspagliesi F, Marcomini B, Canevari S and Mezzanzanica D: Sensitization of p53-mutated epithelial ovarian cancer to CD95-mediated apoptosis is synergistically induced by cisplatin pretreatment. Mol Cancer Ther. 6:762–772. 2007. View Article : Google Scholar : PubMed/NCBI
18	Fulda S and Debatin KM: Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 25:4798–4811. 2006.Bedia C, Casas J, Andrieu-Abadie N, Fabriàs G and Levade T: Acid ceramidase expression modulates the sensitivity of A375 melanoma cells to dacarbazine. J Biol Chem 286: 28200–28209, 2011. View Article : Google Scholar : PubMed/NCBI
19	Bedia C, Casas J, Andrieu-Abadie N, Fabriàs G and Levade T: Acid ceramidase expression modulates the sensitivity of A375 melanoma cells to dacarbazine. J Biol Chem. 286:28200–28209. 2011.Kim YJ, Brox T, Feiden W and Weickert J: Fully automated segmentation and morphometrical analysis of muscle fiber images. Cytometry A 71: 8–15, 2007. View Article : Google Scholar : PubMed/NCBI
20	Kim YJ, Brox T, Feiden W and Weickert J: Fully automated segmentation and morphometrical analysis of muscle fiber images. Cytometry A. 71:8–15. 2007. View Article : Google Scholar : PubMed/NCBI
21	Fulda S and Debatin KM: Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 25:4798–4811. 2006. View Article : Google Scholar : PubMed/NCBI
22	Ghatage DD, Gosavi SR, Ganvir SM and Hazarey VK: Apoptosis: Molecular mechanism. J Orofac Sci. 4:103–107. 2012. View Article : Google Scholar
23	Surova O and Zhivotovsky B: Various modes of cell death induced by DNA damage. Oncogene. 32:3789–3797. 2013. View Article : Google Scholar : PubMed/NCBI
24	Kang MH and Reynolds CP: Bcl-2 inhibitors: Targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res. 15:1126–1132. 2009. View Article : Google Scholar : PubMed/NCBI
25	Yang ZJ, Chee CE, Huang S and Sinicrope FA: The role of autophagy in cancer: Therapeutic implications. Mol Cancer Ther. 10:1533–1534. 2011. View Article : Google Scholar : PubMed/NCBI
26	Tomic T, Botton T, Cerezo M, Robert G, Luciano F, Puissant A, Gounon P, Allegra M, Bertolotto C, Bereder JM, et al: Metformin inhibits melanoma development through autophagy and apoptosis mechanisms. Cell Death Dis. 2:e1992011. View Article : Google Scholar : PubMed/NCBI
27	Zeng Y, Yang X, Wang J, Fan J, Kong Q and Yu X: Aristolochic acid I induced autophagy extenuates cell apoptosis via ERK 1/2 pathway in renal tubular epithelial cells. PLoS One. 7:e303122012. View Article : Google Scholar : PubMed/NCBI
28	Patel PM, Suciu S, Mortier L, Kruit WH, Robert C, Schadendorf D, Trefzer U, Punt CJ, Dummer R, Davidson N, et al: Extended schedule, escalated dose temozolomide versus dacarbazine in stage IV melanoma: Final results of a randomised phase III study (EORTC 18032). Eur J Cancer. 47:1476–1483. 2011. View Article : Google Scholar : PubMed/NCBI
29	Risberg K, Fodstad O and Andersson Y: Anti-melanoma activity of the 9.2.27PE immunotoxin in dacarbazine resistant cells. J Immunother. 33:272–278. 2010. View Article : Google Scholar : PubMed/NCBI
30	Fuller TL and Canada RG: Enhancement of cisplatin cytotoxicity by terbium in cisplatin-resistant MDA/CH human breast cancer cells. Cancer Chemother Pharmacol. 44:249–252. 1999. View Article : Google Scholar : PubMed/NCBI
31	Yang PM and Chen CC: Life or death? Autophagy in anticancer therapies with statins and histone deacetylase inhibitors. Autophagy. 7:107–108. 2011. View Article : Google Scholar : PubMed/NCBI
32	Jin JL, Gong J, Yin TJ, Lu YJ, Xia JJ, Xie YY, Di Y, He L, Guo JL, Sun J, et al: PTD4-apoptin protein and dacarbazine show a synergistic antitumor effect on B16-F1 melanoma in vitro and in vivo. Eur J Pharmacol. 654:17–25. 2011. View Article : Google Scholar : PubMed/NCBI
33	Sanada M, Hidaka M, Takagi Y, Takano TY, Nakatsu Y, Tsuzuki T and Sekiguchi M: Modes of actions of two types of anti-neoplastic drugs, dacarbazine and ACNU, to induce apoptosis. Carcinogenesis. 28:2657–2663. 2007. View Article : Google Scholar : PubMed/NCBI
34	Cun B, Song X, Jia R, Zhao X, Wang H, Ge S and Fan X: Combination of oncolytic adenovirus and dacarbazine enhances antitumor ability against uveal melanoma cells via cell cycle block. Cancer Biol Ther. 13:77–84. 2012. View Article : Google Scholar : PubMed/NCBI
35	Koprowska K, Hartman ML, Sztiller-Sikorska M and Czyz ME: Parthenolide enhances dacarbazine activity against melanoma cells. Cancer Biol. 24:835–845. 2013.
36	Ishibashi M, Arai M, Tanaka S, Onda K and Hirano T: Antiproliferative and apoptosis-inducing effects of lipophilic vitamins on human melanoma A375 cells in vitro. Biol Pharm Bull. 35:10–17. 2012. View Article : Google Scholar : PubMed/NCBI
37	Anvekar RA, Asciolla JJ, Lopez-Rivera E, Floros KV, Izadmehr S, Elkholi R, Belbin G, Sikora AG and Chipuk JE: Sensitization to the mitochondrial pathway of apoptosis augments melanoma tumor cell responses to conventional chemotherapeutic regimens. Cell Death Dis. 3:e4202012. View Article : Google Scholar : PubMed/NCBI
38	Aliwaini S, Peres J, Kröger WL, Blanckenberg A, de la Mare J, Edkins AL, Mapolie S and Prince S: The palladacycle, AJ-5, exhibits anti-tumour and anti-cancer stem cell activity in breast cancer cells. Cancer Lett. 357:206–218. 2015. View Article : Google Scholar : PubMed/NCBI