The epigenetic agents suberoylanilide hydroxamic acid and 5‑AZA‑2' deoxycytidine decrease cell proliferation, induce cell death and delay the growth of MiaPaCa2 pancreatic cancer cells in vivo

Susanto,Johana M.; Colvin,Emily K.; Pinese,Mark; Chang,David K.; Pajic,Marina; Mawson,Amanda; Caldon,C. Elizabeth; Musgrove,Elizabeth A.; Henshall,Susan M.; Sutherland,Robert L.; Biankin,Andrew V.; Scarlett,Christopher J.

doi:10.3892/ijo.2015.2894

The epigenetic agents suberoylanilide hydroxamic acid and 5‑AZA‑2' deoxycytidine decrease cell proliferation, induce cell death and delay the growth of MiaPaCa2 pancreatic cancer cells in vivo

Authors:
- Johana M. Susanto
- Emily K. Colvin
- Mark Pinese
- David K. Chang
- Marina Pajic
- Amanda Mawson
- C. Elizabeth Caldon
- Elizabeth A. Musgrove
- Susan M. Henshall
- Robert L. Sutherland
- Andrew V. Biankin
- Christopher J. Scarlett
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Affiliations: Cancer Research Program, Garvan Institute of Medical Research, Sydney, NSW 2010, Australia
Published online on: February 16, 2015 https://doi.org/10.3892/ijo.2015.2894
Pages: 2223-2230

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Abstract

Despite incremental advances in the diagnosis and treatment for pancreatic cancer (PC), the 5‑year survival rate remains <5%. Novel therapies to increase survival and quality of life for PC patients are desperately needed. Epigenetic therapeutic agents such as histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) have demonstrated therapeutic benefits in human cancer. We assessed the efficacy of these epigenetic therapeutic agents as potential therapies for PC using in vitro and in vivo models. Treatment with HDACi [suberoylanilide hydroxamic acid (SAHA)] and DNMTi [5‑AZA‑2' deoxycytidine (5‑AZA‑dc)] decreased cell proliferation in MiaPaCa2 cells, and SAHA treatment, with or without 5‑AZA‑dc, resulted in higher cell death and lower DNA synthesis compared to 5‑AZA‑dc alone and controls (DMSO). Further, combination treatment with SAHA and 5‑AZA‑dc significantly increased expression of p21WAF1, leading to G1 arrest. Treatment with epigenetic agents delayed tumour growth in vivo, but did not decrease growth of established pancreatic tumours. In conclusion, these data demonstrate a potential role for epigenetic modifier drugs for the management of PC, specifically in the chemoprevention of PC, in combination with other chemotherapeutic agents.

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1	Cancer Institute NSW. Cancer in New South Wales: Incidence, Mortality and Prevalence 2005. Cancer Institute NSW; Sydney: 2007
2	National Cancer Institute: Cancer Trends Progress Report - 2007 Update. National Cancer Institute; Bethesda, MD: 2007
3	National Cancer Institute. A Snapshot of Pancreatic Cancer. National Cancer Institute; Bethesda, MD: 2007
4	Australian Institute of Health and Welfare (AIHW) and Australasian Association of Cancer Registries (AACR). Cancer in Australia: an overview, 2008. AIHW and AACR; Canberra: 2008
5	American Cancer Society. Cancer Facts and Figures 2008. American Cancer Society; Atlanta, GA: 2008
6	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar : PubMed/NCBI
7	American Cancer Society. Cancer Facts and Figures 2007. American Cancer Society; Atlanta, GA: 2007
8	Von Hoff DD, Evans DB and Hruban RH: Pancreatic Cancer. 1st edition. Jones and Bartlett Publishers; Sudbury, MA: 2005
9	Chang DK, Merrett ND and Biankin AV: NSW Pancreatic Cancer Network: Improving outcomes for operable pancreatic cancer: is access to safer surgery the problem? J Gastroenterol Hepatol. 23:1036–1045. 2008. View Article : Google Scholar : PubMed/NCBI
10	Glaser KB: HDAC inhibitors: clinical update and mechanism-based potential. Biochem Pharmacol. 74:659–671. 2007. View Article : Google Scholar : PubMed/NCBI
11	Duvic M, Talpur R, Ni X, et al: Phase 2 trial of oral vorinostat (suberoylanilide hydroxamic acid, SAHA) for refractory cutaneous T-cell lymphoma (CTCL). Blood. 109:31–39. 2007. View Article : Google Scholar
12	Zhang C, Richon V, Ni X, Talpur R and Duvic M: Selective induction of apoptosis by histone deacetylase inhibitor SAHA in cutaneous T-cell lymphoma cells: relevance to mechanism of therapeutic action. J Invest Dermatol. 125:1045–1052. 2005. View Article : Google Scholar : PubMed/NCBI
13	Gore SD: Combination therapy with DNA methyltransferase inhibitors in hematologic malignancies. Nat Clin Pract Oncol. 2(Suppl 1): S30–S35. 2005. View Article : Google Scholar : PubMed/NCBI
14	Issa JP and Byrd JC: Decitabine in chronic leukemias. Semin Hematol. 42(Suppl 2): S43–S49. 2005. View Article : Google Scholar : PubMed/NCBI
15	Batty N, Malouf GG and Issa JP: Histone deacetylase inhibitors as anti-neoplastic agents. Cancer Lett. 280:192–200. 2009. View Article : Google Scholar : PubMed/NCBI
16	Flotho C, Claus R, Batz C, et al: The DNA methyltransferase inhibitors azacitidine, decitabine and zebularine exert differential effects on cancer gene expression in acute myeloid leukemia cells. Leukemia. 23:1019–1028. 2009. View Article : Google Scholar : PubMed/NCBI
17	Laurenzana A, Petruccelli LA, Pettersson F, et al: Inhibition of DNA methyltransferase activates tumor necrosis factor alpha-induced monocytic differentiation in acute myeloid leukemia cells. Cancer Res. 69:55–64. 2009. View Article : Google Scholar : PubMed/NCBI
18	Munster PN, Troso-Sandoval T, Rosen N, Rifkind R, Marks PA and Richon VM: The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces differentiation of human breast cancer cells. Cancer Res. 61:8492–8497. 2001.PubMed/NCBI
19	Shiozawa K, Nakanishi T, Tan M, et al: Preclinical studies of vorinostat (suberoylanilide hydroxamic acid) combined with cytosine arabinoside and etoposide for treatment of acute leukemias. Clin Cancer Res. 15:1698–1707. 2009. View Article : Google Scholar : PubMed/NCBI
20	Xu WS, Parmigiani RB and Marks PA: Histone deacetylase inhibitors: molecular mechanisms of action. Oncogene. 26:5541–5552. 2007. View Article : Google Scholar : PubMed/NCBI
21	Buchwald M, Krämer OH and Heinzel T: HDACi - targets beyond chromatin. Cancer Lett. 280:160–167. 2009. View Article : Google Scholar : PubMed/NCBI
22	Nolan L, Johnson PW, Ganesan A, Packham G and Crabb SJ: Will histone deacetylase inhibitors require combination with other agents to fulfil their therapeutic potential? Br J Cancer. 99:689–694. 2008. View Article : Google Scholar : PubMed/NCBI
23	Gui CY, Ngo L, Xu WS, Richon VM and Marks PA: Histone deacetylase (HDAC) inhibitor activation of p21^WAF1 involves changes in promoter-associated proteins, including HDAC1. Proc Natl Acad Sci USA. 101:1241–1246. 2004. View Article : Google Scholar
24	Arnold NB, Arkus N, Gunn J and Korc M: The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces growth inhibition and enhances gemcitabine-induced cell death in pancreatic cancer. Clin Cancer Res. 13:18–26. 2007. View Article : Google Scholar : PubMed/NCBI
25	Eyüpoglu IY, Hahnen E, Buslei R, et al: Suberoylanilide hydroxamic acid (SAHA) has potent anti-glioma properties in vitro, ex vivo and in vivo. J Neurochem. 93:992–999. 2005. View Article : Google Scholar : PubMed/NCBI
26	Kumagai T, Wakimoto N, Yin D, et al: Histone deacetylase inhibitor, suberoylanilide hydroxamic acid (Vorinostat, SAHA) profoundly inhibits the growth of human pancreatic cancer cells. Int J Cancer. 121:656–665. 2007. View Article : Google Scholar : PubMed/NCBI
27	Hurd PJ, Whitmarsh AJ, Baldwin GS, et al: Mechanism-based inhibition of C5-cytosine DNA methyltransferases by 2-H pyrimidinone. J Mol Biol. 286:389–401. 1999. View Article : Google Scholar : PubMed/NCBI
28	Jones PA and Baylin SB: The fundamental role of epigenetic events in cancer. Nat Rev Genet. 3:415–428. 2002.PubMed/NCBI
29	Jackson-Grusby L, Beard C, Possemato R, et al: Loss of genomic methylation causes p53-dependent apoptosis and epigenetic deregulation. Nat Genet. 27:31–39. 2001. View Article : Google Scholar : PubMed/NCBI
30	Scott SA, Lakshimikuttysamma A, Sheridan DP, Sanche SE, Geyer CR and DeCoteau JF: Zebularine inhibits human acute myeloid leukemia cell growth in vitro in association with p15INK4B demethylation and reexpression. Exp Hematol. 35:263–273. 2007. View Article : Google Scholar : PubMed/NCBI
31	Gilbert J, Gore SD, Herman JG and Carducci MA: The clinical application of targeting cancer through histone acetylation and hypomethylation. Clin Cancer Res. 10:4589–4596. 2004. View Article : Google Scholar : PubMed/NCBI
32	Sato N, Fukushima N, Maehara N, et al: SPARC/osteonectin is a frequent target for aberrant methylation in pancreatic adenocarcinoma and a mediator of tumor-stromal interactions. Oncogene. 22:5021–5030. 2003. View Article : Google Scholar : PubMed/NCBI
33	Sato N, Parker AR, Fukushima N, et al: Epigenetic inactivation of TFPI-2 as a common mechanism associated with growth and invasion of pancreatic ductal adenocarcinoma. Oncogene. 24:850–858. 2005. View Article : Google Scholar
34	Fu B, Guo M, Wang S, et al: Evaluation of GATA-4 and GATA-5 methylation profiles in human pancreatic cancers indicate promoter methylation patterns distinct from other human tumor types. Cancer Biol Ther. 6:1546–1552. 2007. View Article : Google Scholar : PubMed/NCBI
35	Abe T, Toyota M, Suzuki H, et al: Upregulation of BNIP3 by 5-aza-2′-deoxycytidine sensitizes pancreatic cancer cells to hypoxia-mediated cell death. J Gastroenterol. 40:504–510. 2005. View Article : Google Scholar : PubMed/NCBI
36	Yamada N, Nishida Y, Tsutsumida H, et al: MUC1 expression is regulated by DNA methylation and histone H3 lysine 9 modification in cancer cells. Cancer Res. 68:2708–2716. 2008. View Article : Google Scholar : PubMed/NCBI
37	Yamada N, Hamada T, Goto M, et al: MUC2 expression is regulated by histone H3 modification and DNA methylation in pancreatic cancer. Int J Cancer. 119:1850–1857. 2006. View Article : Google Scholar : PubMed/NCBI
38	Vincent A, Ducourouble MP and Van Seuningen I: Epigenetic regulation of the human mucin gene MUC4 in epithelial cancer cell lines involves both DNA methylation and histone modifications mediated by DNA methyltransferases and histone deacetylases. FASEB J. 22:3035–3045. 2008. View Article : Google Scholar : PubMed/NCBI
39	Yonezawa S, Goto M, Yamada N, Higashi M and Nomoto M: Expression profiles of MUC1, MUC2, and MUC4 mucins in human neoplasms and their relationship with biological behavior. Proteomics. 8:3329–3341. 2008. View Article : Google Scholar : PubMed/NCBI
40	Omura N and Goggins M: Epigenetics and epigenetic alterations in pancreatic cancer. Int J Clin Exp Pathol. 2:310–326. 2009.PubMed/NCBI
41	Furukawa T, Duguid WP, Rosenberg L, Viallet J, Galloway DA and Tsao MS: Long-term culture and immortalization of epithelial cells from normal adult human pancreatic ducts transfected by the E6E7 gene of human papilloma virus 16. Am J Pathol. 148:1763–1770. 1996.PubMed/NCBI
42	Vonlaufen A, Joshi S, Qu C, et al: Pancreatic stellate cells: partners in crime with pancreatic cancer cells. Cancer Res. 68:2085–2093. 2008. View Article : Google Scholar : PubMed/NCBI
43	Probst AV, Dunleavy E and Almouzni G: Epigenetic inheritance during the cell cycle. Nat Rev Mol Cell Biol. 10:192–206. 2009. View Article : Google Scholar : PubMed/NCBI
44	Missiaglia E, Donadelli M, Palmieri M, Crnogorac-Jurcevic T, Scarpa A and Lemoine NR: Growth delay of human pancreatic cancer cells by methylase inhibitor 5-aza-2′-deoxycytidine treatment is associated with activation of the interferon signalling pathway. Oncogene. 24:199–211. 2005. View Article : Google Scholar : PubMed/NCBI
45	Bolden JE, Peart MJ and Johnstone RW: Anticancer activities of histone deacetylase inhibitors. Nat Rev Drug Discov. 5:769–784. 2006. View Article : Google Scholar : PubMed/NCBI
46	Butler LM, Agus DB, Scher HI, et al: Suberoylanilide hydroxamic acid, an inhibitor of histone deacetylase, suppresses the growth of prostate cancer cells in vitro and in vivo. Cancer Res. 60:5165–5170. 2000.PubMed/NCBI
47	Butler LM, Zhou X, Xu WS, et al: The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin. Proc Natl Acad Sci USA. 99:11700–11705. 2002. View Article : Google Scholar : PubMed/NCBI
48	Marks PA and Breslow R: Dimethyl sulfoxide to vorinostat: development of this histone deacetylase inhibitor as an anticancer drug. Nat Biotechnol. 25:84–90. 2007. View Article : Google Scholar : PubMed/NCBI
49	Henderson C, Mizzau M, Paroni G, Maestro R, Schneider C and Brancolini C: Role of caspases, Bid, and p53 in the apoptotic response triggered by histone deacetylase inhibitors trichostatin-A (TSA) and suberoylanilide hydroxamic acid (SAHA). J Biol Chem. 278:12579–12589. 2003. View Article : Google Scholar : PubMed/NCBI
50	Egger G, Liang G, Aparicio A and Jones PA: Epigenetics in human disease and prospects for epigenetic therapy. Nature. 429:457–463. 2004. View Article : Google Scholar : PubMed/NCBI
51	Polo SE and Almouzni G: Histone metabolic pathways and chromatin assembly factors as proliferation markers. Cancer Lett. 220:1–9. 2005. View Article : Google Scholar : PubMed/NCBI
52	Koundrioukoff S, Polo S and Almouzni G: Interplay between chromatin and cell cycle checkpoints in the context of ATR/ATM-dependent checkpoints. DNA Repair (Amst). 3:969–978. 2004. View Article : Google Scholar
53	Graham JS, Kaye SB and Brown R: The promises and pitfalls of epigenetic therapies in solid tumours. Eur J Cancer. 45:1129–1136. 2009. View Article : Google Scholar : PubMed/NCBI
54	Polo SE and Almouzni G: DNA damage leaves its mark on chromatin. Cell Cycle. 6:2355–2359. 2007. View Article : Google Scholar : PubMed/NCBI
55	Vrana JA, Decker RH, Johnson CR, et al: Induction of apoptosis in U937 human leukemia cells by suberoylanilide hydroxamic acid (SAHA) proceeds through pathways that are regulated by Bcl-2/Bcl-XL, c-Jun, and p21CIP1, but independent of p53. Oncogene. 18:7016–7025. 1999. View Article : Google Scholar : PubMed/NCBI
56	Abbas T and Dutta A: p21 in cancer: intricate networks and multiple activities. Nat Rev Cancer. 9:400–414. 2009. View Article : Google Scholar : PubMed/NCBI
57	Mills J, Hricik T, Siddiqi S and Matushansky I: Chromatin structure predicts epigenetic therapy responsiveness in sarcoma. Mol Cancer Ther. 10:313–324. 2011. View Article : Google Scholar : PubMed/NCBI
58	Yin D, Ong JM, Hu J, et al: Suberoylanilide hydroxamic acid, a histone deacetylase inhibitor: effects on gene expression and growth of glioma cells in vitro and in vivo. Clin Cancer Res. 13:1045–1052. 2007. View Article : Google Scholar : PubMed/NCBI
59	Vijayaraghavalu S, Dermawan JK, Cheriyath V and Labhasetwar V: Highly synergistic effect of sequential treatment with epigenetic and anticancer drugs to overcome drug resistance in breast cancer cells is mediated via activation of p21 gene expression leading to G2/M cycle arrest. Mol Pharm. 10:337–352. 2013. View Article : Google Scholar :
60	Carew JS, Nawrocki ST and Cleveland JL: Modulating autophagy for therapeutic benefit. Autophagy. 3:464–467. 2007. View Article : Google Scholar : PubMed/NCBI
61	Robert T, Vanoli F, Chiolo I, et al: HDACs link the DNA damage response, processing of double-strand breaks and autophagy. Nature. 471:74–79. 2011. View Article : Google Scholar : PubMed/NCBI
62	Dokmanovic M, Perez G, Xu W, et al: Histone deacetylase inhibitors selectively suppress expression of HDAC7. Mol Cancer Ther. 6:2525–2534. 2007. View Article : Google Scholar : PubMed/NCBI
63	Mitsiades CS, Mitsiades NS, McMullan CJ, et al: Transcriptional signature of histone deacetylase inhibition in multiple myeloma: biological and clinical implications. Proc Natl Acad Sci USA. 101:540–545. 2004. View Article : Google Scholar :
64	Cooper AL, Greenberg VL, Lancaster PS, van Nagell JR Jr, Zimmer SG and Modesitt SC: In vitro and in vivo histone deacetylase inhibitor therapy with suberoylanilide hydroxamic acid (SAHA) and paclitaxel in ovarian cancer. Gynecol Oncol. 104:596–601. 2007. View Article : Google Scholar
65	Nimmanapalli R, Fuino L, Stobaugh C, Richon V and Bhalla K: Cotreatment with the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) enhances imatinib-induced apoptosis of Bcr-Abl-positive human acute leukemia cells. Blood. 101:3236–3239. 2003. View Article : Google Scholar
66	Holzman DC: Pancreatic cancer: will incremental advances begin to make a difference? J Natl Cancer Inst. 102:1821–1823. 2010. View Article : Google Scholar : PubMed/NCBI
67	Neoptolemos JP, Dunn JA, Stocken DD, et al: Adjuvant chemoradiotherapy and chemotherapy in resectable pancreatic cancer: a randomised controlled trial. Lancet. 358:1576–1585. 2001. View Article : Google Scholar : PubMed/NCBI
68	Neoptolemos JP, Stocken DD, Bassi C, et al: Adjuvant chemotherapy with fluorouracil plus folinic acid vs gemcitabine following pancreatic cancer resection: a randomized controlled trial. JAMA. 304:1073–1081. 2010. View Article : Google Scholar : PubMed/NCBI
69	O’Reilly EM: Refinement of adjuvant therapy for pancreatic cancer. JAMA. 304:1124–1125. 2010. View Article : Google Scholar
70	Mohammed A, Janakiram NB, Li Q, et al: The epidermal growth factor receptor inhibitor gefitinib prevents the progression of pancreatic lesions to carcinoma in a conditional LSL-KrasG12D/+ transgenic mouse model. Cancer Prev Res (Phila). 3:1417–1426. 2010. View Article : Google Scholar
71	Appleton K, Mackay HJ, Judson I, et al: Phase I and pharmacodynamic trial of the DNA methyltransferase inhibitor decitabine and carboplatin in solid tumors. J Clin Oncol. 25:4603–4609. 2007. View Article : Google Scholar : PubMed/NCBI
72	Venturelli S, Berger A, Weiland T, et al: Dual antitumour effect of 5-azacytidine by inducing a breakdown of resistance-mediating factors and epigenetic modulation. Gut. 60:156–165. 2011. View Article : Google Scholar
73	Shakya R, Gonda T, Quante M, et al: Hypomethylating therapy in an aggressive stroma-rich model of pancreatic carcinoma. Cancer Res. 73:885–896. 2013. View Article : Google Scholar :