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Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2014

Open Access 01-12-2014 | Research

The inhibitory effects of extracellular ATP on the growth of nasopharyngeal carcinoma cells via P2Y2 receptor and osteopontin

Authors: Guang Yang, Shenghong Zhang, Yanling Zhang, Qiming Zhou, Sheng Peng, Tao Zhang, Changfu Yang, Zhenyu Zhu, Fujun Zhang

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2014

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Abstract

Background

Nasopharyngeal carcinoma (NPC) is a common malignant tumor observed in the populations of southern China and Southeast Asia. However, little is known about the effects of purinergic signal on the behavior of NPC cells. This study analyzed the effects of ATP on the growth and migration of NPC cells, and further investigated the potential mechanisms during the effects.

Methods

Cell viability was estimated by MTT assay. Transwell assay was utilized to assess the motility of NPC cells. Cell cycle and apoptosis were detected by flow cytometry analysis. Changes in OPN, P2Y2 and p65 expression were assessed by western blotting analysis or immunofluorescence. The effects of ATP and P2Y2 on promoter activity of OPN were analyzed by luciferase activity assay. The binding of p65 to the promoter region of OPN was examined by ChIP assay.

Results

An MTT assay indicated that ATP inhibited the proliferation of NPC cells in time- and dose-dependent manners, and a Transwell assay showed that extracellular ATP inhibited the motility of NPC cells. We further investigated the potential mechanisms involved in the inhibitory effect of extracellular ATP on the growth of NPC cells and found that extracellular ATP could reduce Bcl-2 and p-AKT levels while elevating Bax and cleaved caspase-3 levels in NPC cells. Decreased levels of p65 and osteopontin were also detected in the ATP-treated NPC cells. We demonstrated that extracellular ATP inhibited the growth of NPC cells via p65 and osteopontin and verified that P2Y2 overexpression elevated the inhibitory effect of extracellular ATP on the proliferation of NPC cells. Moreover, a dual luciferase reporter assay showed that the level of osteopontin transcription was inhibited by extracellular ATP and P2Y2. ATP decreased the binding of p65 to potential sites in the OPN promoter region in NPC cells.

Conclusion

This study indicated that extracellular ATP inhibited the growth of NPC cells via P2Y2, p65 and OPN. ATP could be a promising agent serving as an adjuvant in the treatment of NPC.
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Literature
1.
2.
Ralevic V, Burnstock G: Receptors for purines and pyrimidines. Pharmacol Rev. 1998, 50 (3): 413-492.PubMed
3.
Yaguchi T, Saito M, Yasuda Y, Kanno T, Nakano T, Nishizaki T: Higher concentrations of extracellular ATP suppress proliferation of Caco-2 human colonic cancer cells via an unknown receptor involving PKC inhibition. Cell Physiol Biochem. 2010, 26 (2): 125-134.CrossRefPubMed
4.
Maaser K, Hopfner M, Kap H, Sutter AP, Barthel B, von Lampe B, Zeitz M, Scherubl H: Extracellular nucleotides inhibit growth of human oesophageal cancer cells via P2Y(2)-receptors. Br J Cancer. 2002, 86 (4): 636-644.PubMedCentralCrossRefPubMed
5.
6.
Volkl T, Ogilvie A, Neuhuber W, Ogilvie A: Cell death induced by uridine 5′-triphosphate (UTP) in contrast to adenosine 5′-triphosphate (ATP) in human epidermoid carcinoma cells (A-431). Cell Physiol Biochem. 2008, 22 (5–6): 441-454.PubMed
7.
Abbracchio MP, Burnstock G: Purinoceptors: are there families of P2X and P2Y purinoceptors?. Pharmacol Ther. 1994, 64 (3): 445-475.CrossRefPubMed
8.
9.
Sak K, Webb TE: A retrospective of recombinant P2Y receptor subtypes and their pharmacology. Arch Biochem Biophys. 2002, 397 (1): 131-136.CrossRefPubMed
10.
Khakh BS, Burnstock G, Kennedy C, King BF, North RA, Seguela P, Voigt M, Humphrey PP: International union of pharmacology. XXIV. Current status of the nomenclature and properties of P2X receptors and their subunits. Pharmacol Rev. 2001, 53 (1): 107-118.PubMed
11.
Dubyak GR, El-Moatassim C: Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol. 1993, 265 (3 Pt 1): C577-C606.PubMed
12.
Faure E, Garrouste F, Parat F, Monferran S, Leloup L, Pommier G, Kovacic H, Lehmann M: P2Y2 receptor inhibits EGF-induced MAPK pathway to stabilise keratinocyte hemidesmosomes. J Cell Sci. 2012, 125 (Pt 18): 4264-4277.CrossRefPubMed
13.
Seye CI, Agca Y, Agca C, Derbigny W: P2Y2 receptor-mediated lymphotoxin-alpha secretion regulates intercellular cell adhesion molecule-1 expression in vascular smooth muscle cells. J Biol Chem. 2012, 287 (13): 10535-10543.PubMedCentralCrossRefPubMed
14.
Arthur DB, Akassoglou K, Insel PA: P2Y2 receptor activates nerve growth factor/TrkA signaling to enhance neuronal differentiation. Proc Natl Acad Sci USA. 2005, 102 (52): 19138-19143.PubMedCentralCrossRefPubMed
15.
Elliott MR, Chekeni FB, Trampont PC, Lazarowski ER, Kadl A, Walk SF, Park D, Woodson RI, Ostankovich M, Sharma P, Lysiak JJ, Harden TK, Leitinger N, Ravichandran KS: Nucleotides released by apoptotic cells act as a find-me signal to promote phagocytic clearance. Nature. 2009, 461 (7261): 282-286.PubMedCentralCrossRefPubMed
16.
Chen Y, Liu H, Wu W, Li Y, Li J: Osteopontin genetic variants are associated with overall survival in advanced non-small-cell lung cancer patients and bone metastasis. J Exp Clin Cancer Res. 2013, 32: 45-PubMedCentralCrossRefPubMed
17.
Shojaei F, Scott N, Kang X, Lappin PB, Fitzgerald AA, Karlicek S, Simmons BH, Wu A, Lee JH, Bergqvist S, Kraynov E: Osteopontin induces growth of metastatic tumors in a preclinical model of non-small lung cancer. J Exp Clin Cancer Res. 2012, 31: 26-PubMedCentralCrossRefPubMed
18.
Shang S, Plymoth A, Ge S, Feng Z, Rosen HR, Sangrajrang S, Hainaut P, Marrero JA, Beretta L: Identification of osteopontin as a novel marker for early hepatocellular carcinoma. Hepatology. 2012, 55 (2): 483-490.PubMedCentralCrossRefPubMed
19.
Sun BS, Dong QZ, Ye QH, Sun HJ, Jia HL, Zhu XQ, Liu DY, Chen J, Xue Q, Zhou HJ, Ren N, Qin LX: Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma. Hepatology. 2008, 48 (6): 1834-1842.CrossRefPubMed
20.
Lin YH, Yang-Yen HF: The osteopontin-CD44 survival signal involves activation of the phosphatidylinositol 3-kinase/Akt signaling pathway. J Biol Chem. 2001, 276 (49): 46024-46030.CrossRefPubMed
21.
Jalvy S, Renault MA, Lam SLL, Belloc I, Reynaud A, Gadeau AP, Desgranges C: CREB mediates UTP-directed arterial smooth muscle cell migration and expression of the chemotactic protein osteopontin via its interaction with activator protein-1 sites. Circ Res. 2007, 100 (9): 1292-1299.CrossRefPubMed
22.
Cheng J, Huo DH, Kuang DM, Yang J, Zheng L, Zhuang SM: Human macrophages promote the motility and invasiveness of osteopontin-knockdown tumor cells. Cancer Res. 2007, 67 (11): 5141-5147.CrossRefPubMed
23.
Yang G, Zhang Y, Wu J, Xiong J, Deng H, Wang J, Yang C, Zhu Z: Osteopontin regulates growth and migration of human nasopharyngeal cancer cells. Mol Med Rep. 2011, 4 (6): 1169-1173.PubMed
24.
Kim MS, Park MJ, Moon EJ, Kim SJ, Lee CH, Yoo H, Shin SH, Song ES, Lee SH: Hyaluronic acid induces osteopontin via the phosphatidylinositol 3-kinase/Akt pathway to enhance the motility of human glioma cells. Cancer Res. 2005, 65 (3): 686-691.PubMed
25.
Zhang G, He B, Weber GF: Growth factor signaling induces metastasis genes in transformed cells: molecular connection between Akt kinase and osteopontin in breast cancer. Mol Cell Biol. 2003, 23 (18): 6507-6519.PubMedCentralCrossRefPubMed
26.
Chen HT, Tsou HK, Chang CH, Tang CH: Hepatocyte growth factor increases osteopontin expression in human osteoblasts through PI3K, Akt, c-Src, and AP-1 signaling pathway. PLoS One. 2012, 7 (6): e38378-PubMedCentralCrossRefPubMed
27.
Frazzi R, Valli R, Tamagnini I, Casali B, Latruffe N, Merli F: Resveratrol-mediated apoptosis of hodgkin lymphoma cells involves SIRT1 inhibition and FOXO3a hyperacetylation. Int J Cancer. 2013, 132 (5): 1013-1021.CrossRefPubMed
28.
Kolfschoten GM, Hulscher TM, Pinedo HM, Boven E: Drug resistance features and S-phase fraction as possible determinants for drug response in a panel of human ovarian cancer xenografts. Br J Cancer. 2000, 83 (7): 921-927.PubMedCentralCrossRefPubMed
29.
Wagstaff SC, Bowler WB, Gallagher JA, Hipskind RA: Extracellular ATP activates multiple signalling pathways and potentiates growth factor-induced c-fos gene expression in MCF-7 breast cancer cells. Carcinogenesis. 2000, 21 (12): 2175-2181.CrossRefPubMed
30.
Nandigama R, Padmasekar M, Wartenberg M, Sauer H: Feed forward cycle of hypotonic stress-induced ATP release, purinergic receptor activation, and growth stimulation of prostate cancer cells. J Biol Chem. 2006, 281 (9): 5686-5693.CrossRefPubMed
31.
Mao J, Wang L, Fan A, Wang J, Xu B, Jacob TJ, Chen L: Blockage of volume-activated chloride channels inhibits migration of nasopharyngeal carcinoma cells. Cell Physiol Biochem. 2007, 19 (5–6): 249-258.CrossRefPubMed
32.
Packer L, Pavey S, Parker A, Stark M, Johansson P, Clarke B, Pollock P, Ringner M, Hayward N: Osteopontin is a downstream effector of the PI3-kinase pathway in melanomas that is inversely correlated with functional PTEN. Carcinogenesis. 2006, 27 (9): 1778-1786.CrossRefPubMed
33.
Wong TS, Kwong DL, Sham J, Wei WI, Kwong YL, Yuen AP: Elevation of plasma osteopontin level in patients with undifferentiated nasopharyngeal carcinoma. Eur J Surg Oncol. 2005, 31 (5): 555-558.CrossRefPubMed
34.
Hui EP, Sung FL, Yu BK, Wong CS, Ma BB, Lin X, Chan A, Wong WL, Chan AT: Plasma osteopontin, hypoxia, and response to radiotherapy in nasopharyngeal cancer. Clin Cancer Res. 2008, 14 (21): 7080-7087.CrossRefPubMed
35.
Crooke A, Mediero A, Guzman-Aranguez A, Pintor J: Silencing of P2Y2 receptor delays Ap4A-corneal re-epithelialization process. Mol Vis. 2009, 15: 1169-1178.PubMedCentralPubMed
36.
Taboubi S, Milanini J, Delamarre E, Parat F, Garrouste F, Pommier G, Takasaki J, Hubaud JC, Kovacic H, Lehmann M: G alpha(q/11)-coupled P2Y2 nucleotide receptor inhibits human keratinocyte spreading and migration. FASEB J. 2007, 21 (14): 4047-4058.CrossRefPubMed
37.
Karin M: Nuclear factor-kappaB in cancer development and progression. Nature. 2006, 441 (7092): 431-436.CrossRefPubMed
38.
Hann SS, Zheng F, Zhao S: Targeting 3-phosphoinositide-dependent protein kinase 1 by N-acetyl-cysteine through activation of peroxisome proliferators activated receptor alpha in human lung cancer cells, the role of p53 and p65. J Exp Clin Cancer Res. 2013, 32: 43-PubMedCentralCrossRefPubMed
39.
Watari K, Nakamura M, Fukunaga Y, Furuno A, Shibata T, Kawahara A, Hosoi F, Kuwano T, Kuwano M, Ono M: The antitumor effect of a novel angiogenesis inhibitor (an octahydronaphthalene derivative) targeting both VEGF receptor and NF-kappaB pathway. Int J Cancer. 2012, 131 (2): 310-321.CrossRefPubMed
40.
Xiao Z, Yang M, Fang L, Lv Q, He Q, Deng M, Liu X, Chen X, Chen M, Xie X, Hu J: Extracellular nucleotide inhibits cell proliferation and negatively regulates Toll-like receptor 4 signalling in human progenitor endothelial cells. Cell Biol Int. 2012, 36 (7): 625-633.CrossRefPubMed
Metadata
Title
The inhibitory effects of extracellular ATP on the growth of nasopharyngeal carcinoma cells via P2Y2 receptor and osteopontin
Authors
Guang Yang
Shenghong Zhang
Yanling Zhang
Qiming Zhou
Sheng Peng
Tao Zhang
Changfu Yang
Zhenyu Zhu
Fujun Zhang
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2014
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/1756-9966-33-53

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