DFMO/eflornithine inhibits migration and invasion downstream of MYCN and involves p27Kip1 activity in neuroblastoma

Koomoa,Dana-Lynn T.; Geerts,Dirk; Lange,Ingo; Koster,Jan; Pegg,Anthony E.; Feith,David J.; Bachmann,André S.

doi:10.3892/ijo.2013.1835

DFMO/eflornithine inhibits migration and invasion downstream of MYCN and involves p27Kip1 activity in neuroblastoma

Authors:
- Dana-Lynn T. Koomoa
- Dirk Geerts
- Ingo Lange
- Jan Koster
- Anthony E. Pegg
- David J. Feith
- André S. Bachmann
View Affiliations

Affiliations: Department of Pharmaceutical Sciences, College of Pharmacy, University of Hawaii at Hilo, Hilo, HI 96720, USA, Department of Pediatric Oncology/Hematology, Sophia Children's Hospital, Erasmus University Medical Center, 3015 GE Rotterdam, The Netherlands, Department of Human Genetics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Published online on: February 21, 2013 https://doi.org/10.3892/ijo.2013.1835
Pages: 1219-1228

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Neuroblastoma (NB) is the most common extracranial pediatric tumor. NB patients over 18 months of age at the time of diagnosis are often in the later stages of the disease, present with widespread dissemination, and often possess MYCN tumor gene amplification. MYCN is a transcription factor that regulates the expression of a number of genes including ornithine decarboxylase (ODC), a rate-limiting enzyme in the biosynthesis of polyamines. Inhibiting ODC in NB cells produces many deleterious effects including G1 cell cycle arrest, inhibition of cell proliferation, and decreased tumor growth, making ODC a promising target for drug interference. DFMO treatment leads to the accumulation of the cyclin-dependent kinase inhibitor p27Kip1 protein and causes p27Kip1/Rb-coupled G1 cell cycle arrest in MYCN-amplified NB tumor cells through a process that involves p27Kip1 phosphorylation at residues Ser10 and Thr198. While p27Kip1 is well known for its role as a cyclin-dependent kinase inhibitor, recent studies have revealed a novel function of p27Kip1 as a regulator of cell migration and invasion. In the present study we found that p27Kip1 regulates the migration and invasion in NB and that these events are dependent on the state of phosphorylation of p27Kip1. DFMO treatments induced MYCN protein downregulation and phosphorylation of Akt/PKB (Ser473) and GSK3-β (Ser9), and polyamine supplementation alleviated the DFMO-induced effects. Importantly, we provide strong evidence that p27Kip1 mRNA correlates with clinical features and the survival probability of NB patients.

View Figures

View References

1	Maris JM, Hogarty MD, Bagatell R and Cohn SL: Neuroblastoma. Lancet. 369:2106–2120. 2007. View Article : Google Scholar : PubMed/NCBI
2	Maris JM: Recent advances in neuroblastoma. N Engl J Med. 362:2202–2211. 2010. View Article : Google Scholar : PubMed/NCBI
3	Schwab M, Westermann F, Hero B and Berthold F: Neuroblastoma: biology and molecular and chromosomal pathology. Lancet Oncol. 4:472–480. 2003. View Article : Google Scholar : PubMed/NCBI
4	Brodeur GM: Neuroblastoma: biological insights into a clinical enigma. Nat Rev Cancer. 3:203–216. 2003. View Article : Google Scholar : PubMed/NCBI
5	Canete A, Gerrard M, Rubie H, et al: Poor survival for infants with MYCN-amplified metastatic neuroblastoma despite intensified treatment: the International Society of Paediatric Oncology European Neuroblastoma Experience. J Clin Oncol. 27:1014–1019. 2009. View Article : Google Scholar
6	De Bernardi B, Gerrard M, Boni L, et al: Excellent outcome with reduced treatment for infants with disseminated neuroblastoma without MYCN gene amplification. J Clin Oncol. 27:1034–1040. 2009.PubMed/NCBI
7	Johnsen JI, Segerstrom L, Orrego A, et al: Inhibitors of mammalian target of rapamycin downregulate MYCN protein expression and inhibit neuroblastoma growth in vitro and in vivo. Oncogene. 27:2910–2922. 2008. View Article : Google Scholar : PubMed/NCBI
8	Otto T, Horn S, Brockmann M, et al: Stabilization of N-Myc is a critical function of Aurora A in human neuroblastoma. Cancer Cell. 15:67–78. 2009. View Article : Google Scholar : PubMed/NCBI
9	Ushmorov A, Hogarty MD, Liu X, Knauss H, Debatin KM and Beltinger C: N-myc augments death and attenuates protective effects of Bcl-2 in trophically stressed neuroblastoma cells. Oncogene. 27:3424–3434. 2008. View Article : Google Scholar : PubMed/NCBI
10	Wallick CJ, Gamper I, Thorne M, et al: Key role for p27^Kip1, retinoblastoma protein Rb, and MYCN in polyamine inhibitor-induced G1 cell cycle arrest in MYCN-amplified human neuroblastoma cells. Oncogene. 24:5606–5618. 2005.PubMed/NCBI
11	Koomoa DL, Yco LP, Borsics T, Wallick CJ and Bachmann AS: Ornithine decarboxylase inhibition by alpha-difluoromethylornithine activates opposing signaling pathways via phosphorylation of both Akt/protein kinase B and p27^Kip1 in neuroblastoma. Cancer Res. 68:9825–9831. 2008. View Article : Google Scholar
12	Chiarle R, Pagano M and Inghirami G: The cyclin dependent kinase inhibitor p27 and its prognostic role in breast cancer. Breast Cancer Res. 3:91–94. 2001. View Article : Google Scholar : PubMed/NCBI
13	Chu IM, Hengst L and Slingerland JM: The Cdk inhibitor p27 in human cancer: prognostic potential and relevance to anticancer therapy. Nat Rev Cancer. 8:253–267. 2008. View Article : Google Scholar : PubMed/NCBI
14	Esposito V, Baldi A, De Luca A, et al: Prognostic role of the cyclin-dependent kinase inhibitor p27 in non-small cell lung cancer. Cancer Res. 57:3381–3385. 1997.PubMed/NCBI
15	Lee J and Kim SS: The function of p27^Kip1 during tumor development. Exp Mol Med. 41:765–771. 2009.
16	Ravanko K, Jarvinen K, Paasinen-Sohns A and Holtta E: Loss of p27^Kip1 from cyclin E/cyclin-dependent kinase (CDK) 2 but not from cyclin D1/CDK4 complexes in cells transformed by polyamine biosynthetic enzymes. Cancer Res. 60:5244–5253. 2000.PubMed/NCBI
17	Baldassarre G, Belletti B, Nicoloso MS, et al: p27(Kip1)-stathmin interaction influences sarcoma cell migration and invasion. Cancer Cell. 7:51–63. 2005. View Article : Google Scholar : PubMed/NCBI
18	Itoh Y, Masuyama N, Nakayama K, Nakayama KI and Gotoh Y: The cyclin-dependent kinase inhibitors p57 and p27 regulate neuronal migration in the developing mouse neocortex. J Biol Chem. 282:390–396. 2007. View Article : Google Scholar : PubMed/NCBI
19	Li Z, Jiao X, Wang C, et al: Cyclin D1 induction of cellular migration requires p27(^Kip1). Cancer Res. 66:9986–9994. 2006. View Article : Google Scholar : PubMed/NCBI
20	McAllister SS, Becker-Hapak M, Pintucci G, Pagano M and Dowdy SF: Novel p27(^Kip1) C-terminal scatter domain mediates Rac-dependent cell migration independent of cell cycle arrest functions. Mol Cell Biol. 23:216–228. 2003.
21	See WL, Heinberg AR, Holland EC and Resh MD: p27 deficiency is associated with migration defects in PDGF-expressing gliomas in vivo. Cell Cycle. 9:1562–1567. 2010. View Article : Google Scholar : PubMed/NCBI
22	Stehr W, Mercer TI, Bernal NP, Erwin CR and Warner BW: Opposing roles for p21(waf1/cip1) and p27(^Kip1) in enterocyte differentiation, proliferation, and migration. Surgery. 138:187–194. 2005.PubMed/NCBI
23	Sun J, Marx SO, Chen HJ, Poon M, Marks AR and Rabbani LE: Role for p27(Kip1) in vascular smooth muscle cell migration. Circulation. 103:2967–2972. 2001. View Article : Google Scholar : PubMed/NCBI
24	Woods TC: Regulation of cell migration by mTOR is mediated through changes in p27(Kip1) phosphorylation. Cell Cycle. 9:2057–2058. 2010. View Article : Google Scholar : PubMed/NCBI
25	Assoian RK: Stopping and going with p27^Kip1. Dev Cell. 6:458–459. 2004. View Article : Google Scholar : PubMed/NCBI
26	Besson A, Gurian-West M, Schmidt A, Hall A and Roberts JM: p27^Kip1 modulates cell migration through the regulation of RhoA activation. Genes Dev. 18:862–876. 2004.
27	Larrea MD, Wander SA and Slingerland JM: p27 as Jekyll and Hyde: regulation of cell cycle and cell motility. Cell Cycle. 8:3455–3461. 2009. View Article : Google Scholar : PubMed/NCBI
28	Iancu-Rubin C and Atweh GF: p27(^Kip1) and stathmin share the stage for the first time. Trends Cell Biol. 15:346–348. 2005.
29	Denicourt C, Saenz CC, Datnow B, Cui XS and Dowdy SF: Relocalized p27^Kip1 tumor suppressor functions as a cytoplasmic metastatic oncogene in melanoma. Cancer Res. 67:9238–9243. 2007.PubMed/NCBI
30	Kossatz U, Vervoorts J, Nickeleit I, et al: C-terminal phosphorylation controls the stability and function of p27^Kip1. EMBO J. 25:5159–5170. 2006. View Article : Google Scholar : PubMed/NCBI
31	Sicinski P, Zacharek S and Kim C: Duality of p27^Kip1 function in tumorigenesis. Genes Dev. 21:1703–1706. 2007. View Article : Google Scholar
32	Supriatno, Harada K, Kawaguchi S, Yoshida H and Sato M: Effect of p27^Kip1 on the ability of invasion and metastasis of an oral cancer cell line. Oncol Rep. 10:527–532. 2003.
33	Wang XQ, Lui EL, Cai Q, et al: p27^Kip1 promotes migration of metastatic hepatocellular carcinoma cells. Tumour Biol. 29:217–223. 2008.
34	Geerts D, Koster J, Albert D, et al: The polyamine metabolism genes ornithine decarboxylase and antizyme 2 predict aggressive behavior in neuroblastomas with and without MYCN amplification. Int J Cancer. 126:2012–2024. 2010.
35	Koomoa DL, Borsics T, Feith DJ, et al: Inhibition of S-adenosylmethionine decarboxylase by inhibitor SAM486A connects polyamine metabolism with p53-Mdm2-Akt/protein kinase B regulation and apoptosis in neuroblastoma. Mol Cancer Ther. 8:2067–2075. 2009. View Article : Google Scholar
36	Revet I, Huizenga G, Chan A, et al: The MSX1 homeobox transcription factor is a downstream target of PHOX2B and activates the Delta-Notch pathway in neuroblastoma. Exp Cell Res. 314:707–719. 2008. View Article : Google Scholar : PubMed/NCBI
37	Motti ML, Califano D, Troncone G, et al: Complex regulation of the cyclin-dependent kinase inhibitor p27^Kip1 in thyroid cancer cells by the PI3K/AKT pathway: regulation of p27^Kip1 expression and localization. Am J Pathol. 166:737–749. 2005. View Article : Google Scholar : PubMed/NCBI
38	Motti ML, De Marco C, Califano D, Fusco A and Viglietto G: Akt-dependent T198 phosphorylation of cyclin-dependent kinase inhibitor p27^Kip1 in breast cancer. Cell Cycle. 3:1074–1080. 2004. View Article : Google Scholar : PubMed/NCBI
39	van Weeren PC, de Bruyn KM, de Vries-Smits AM, van Lint J and Burgering BM: Essential role for protein kinase B (PKB) in insulin-induced glycogen synthase kinase 3 inactivation. Characterization of dominant-negative mutant of PKB. J Biol Chem. 273:13150–13156. 1998.PubMed/NCBI
40	Bergmann E, Wanzel M, Weber A, Shin I, Christiansen H and Eilers M: Expression of P27(^Kip1) is prognostic and independent of MYCN amplification in human neuroblastoma. Int J Cancer. 95:176–183. 2001.
41	Borriello A, Bencivenga D, Criscuolo M, et al: Targeting p27(^Kip1) protein: its relevance in the therapy of human cancer. Expert Opin Ther Targets. 15:677–693. 2011.
42	Richert MM, Phadke PA, Matters G, et al: Metastasis of hormone-independent breast cancer to lung and bone is decreased by alpha-difluoromethylornithine treatment. Breast Cancer Res. 7:R819–R827. 2005. View Article : Google Scholar
43	Manni A, Washington S, Craig L, et al: Effects of alpha-difluoromethylornithine on local recurrence and pulmonary metastasis from MDA-MB-435 breast cancer xenografts in nude mice. Clin Exp Metastasis. 20:321–325. 2003. View Article : Google Scholar : PubMed/NCBI
44	Jun JY, Griffith JW, Bruggeman R, et al: Effects of polyamine depletion by alpha-difluoromethylornithine on in vitro and in vivo biological properties of 4T1 murine mammary cancer cells. Breast Cancer Res Treat. 105:29–36. 2007. View Article : Google Scholar
45	Ohmori T, Okada K, Tabei R and Shibata T: Effects on tumor induction, growth, metastasis and histology of concurrent administration of putrescine and its metabolizing inhibitor alpha-difluoromethylornithine in nickel tumorigenesis in soft tissue. Carcinogenesis. 15:647–652. 1994. View Article : Google Scholar
46	Zirvi KA, Dasmahapatra KS, Atabek U and Lyons MA: alpha-Difluoromethylornithine inhibits liver metastasis produced by intrasplenic injection of human tumor cells into nude mice. Clin Exp Metastasis. 7:591–598. 1989. View Article : Google Scholar
47	Kubota S, Ohsawa N and Takaku F: Effects of DL-alpha-difluoromethylornithine on the growth and metastasis of B16 melanoma in vivo. Int J Cancer. 39:244–247. 1987. View Article : Google Scholar : PubMed/NCBI
48	Sunkara PS and Rosenberger AL: Antimetastatic activity of DL-alpha-difluoromethylornithine, an inhibitor of polyamine biosynthesis, in mice. Cancer Res. 47:933–935. 1987.PubMed/NCBI
49	Meyskens FL, Kingsley EM, Glattke T, Loescher L and Booth A: A phase II study of alpha-difluoromethylornithine (DFMO) for the treatment of metastatic melanoma. Invest New Drugs. 4:257–262. 1986. View Article : Google Scholar : PubMed/NCBI
50	Klein S, Miret JJ, Algranati ID and de Lustig ES: Effect of alpha-difluoromethylornithine in lung metastases before and after surgery of primary adenocarcinoma tumors in mice. Biol Cell. 53:33–36. 1985. View Article : Google Scholar : PubMed/NCBI
51	Yaari S, Jacob-Hirsch J, Amariglio N, Haklai R, Rechavi G and Kloog Y: Disruption of cooperation between Ras and MycN in human neuroblastoma cells promotes growth arrest. Clin Cancer Res. 11:4321–4330. 2005. View Article : Google Scholar : PubMed/NCBI
52	Hogarty MD, Norris MD, Davis K, et al: ODC1 is a critical determinant of MYCN oncogenesis and a therapeutic target in neuroblastoma. Cancer Res. 68:9735–9745. 2008. View Article : Google Scholar : PubMed/NCBI
53	Rounbehler RJ, Li W, Hall MA, Yang C, Fallahi M and Cleveland JL: Targeting ornithine decarboxylase impairs development of MYCN-amplified neuroblastoma. Cancer Res. 69:547–553. 2009. View Article : Google Scholar : PubMed/NCBI