Potential role of P2X7R in esophageal squamous cell carcinoma proliferation
- 372 Downloads
Esophageal cancer is an aggressive tumor and is the sixth leading cause of cancer death worldwide. ATP is well known to regulate cancer progression in a variety of models by different mechanisms, including P2X7R activation. This study aimed to evaluate the role of P2X7R in esophageal squamous cell carcinoma (ESCC) proliferation. Our results show that treatment with high ATP concentrations induced a decrease in cell number, cell viability, number of polyclonal colonies, and reduced migration of ESCC. The treatment with the selective P2X7R antagonist A740003 or siRNA for P2X7 reverted this effect in the KYSE450 cell line. In addition, results showed that P2X7R is highly expressed, at mRNA and protein levels, in KYSE450 lineage. Additionally, KYSE450, KYSE30, and OE21 cells express P2X3R, P2X4R, P2X5R, P2X6R, and P2X7R genes. P2X1R is expressed by KYSE30 and KYSE450, and only KYSE450 expresses the P2X2R gene. Furthermore, esophageal cancer cell line KYSE450 presented higher expression of E-NTPDases 1 and 2 and of Ecto-5′-NT/CD73 when compared to normal cells. This cell line also exhibits ATPase, ADPase, and AMPase activity, although in different levels, and the co-treatment of apyrase was able to revert the antiproliferative effects of ATP. Moreover, results showed high immunostaining for P2X7R in biopsies of patients with esophageal carcinoma, indicating the involvement of this receptor in the growth of this type of cancer. The results suggest that P2X7R may be a potential pharmacological target to treat ESCC and can lead us to further investigate the effect of this receptor in cancer cell progression.
KeywordsP2X7R Esophageal cancer Proliferation ATP Ectonucleotidases
This work was supported by CAPES and CNPq scholarships, PUCRS, and FINEP research grant “Implantação, Modernização e Qualificação de Estrutura de Pesquisa da PUCRS” (PUCRSINFRA) no. 01.11.0014-00. We would like to thank Dr. Guido Lenz from UFRGS for P2X7R antibody donation, Dr. Mohamed I Parker from ICGEB, Dr. Ana Maria O. Battastini from UFRGS, and Dr. Maria Martha Campos from PUCRS for their intellectual and technical advice.
Compliance with ethical standards
Conflicts of interest
André A Santos Jr declares that he has no conflict of interest.
Angélica R Cappellari declares that she has no conflict of interest.
Fernanda O de Marchi declares that she has no conflict of interest.
Marina P Gehring declares that she has no conflict of interest.
Aline Zaparte declares that she has no conflict of interest.
Caroline A Brandão declares that she has no conflict of interest.
Tiago Giuliani Lopes declares that he has no conflict of interest.
Luis Felipe Ribeiro Pinto that he has no conflict of interest.
Vinicius Duval da Silva declares that he has no conflict of interest.
Robson Coutinho-Silva declares that he has no conflict of interest.
Juliano D Paccez declares that she has no conflict of interest.
Luiz F Zerbini declares that he has no conflict of interest.
Fernanda B Morrone declares that she has no conflict of interest.
Histological samples of human ESCC and esophageal tissue samples of patients with esophagitis were collected, between July and December 2015, from patients who underwent endoscopic procedures with biopsies and/or surgical resection at Pontificia Universidade Católica do Rio Grande do Sul (PUCRS, Porto Alegre, Brazil). The diagnosis was reviewed by two certified pathologists with at least 20-year experience in surgical pathology. Samples were obtained in accordance with approved ethical standards of the Institutional Research Ethics Committee (CAAE 4969 6115.0.0000.5336).
- 2.Instituto Nacional de Câncer José de Alencar Gomes da Silva (2015) Estimate (2016) - Cancer Incidence in Brazil. Rio de Janeiro, INCAGoogle Scholar
- 4.Morita M, Kumashiro R, Kubo N, Nakashima Y, Yoshida R, Yoshinaga K, Saeki H, Emi Y, KakejiY SY, Toh Y, Maehara Y (2010) Alcohol drinking, cigarette smoking, and the development of squamous cell carcinoma of the esophagus: epidemiology, clinical findings, and prevention. Int J ClinOncol 15(2):126–134. doi: 10.1007/s10147-010-0056-7 Google Scholar
- 5.Verschuur EM, Siersema PD (2010) Diagnostics and treatment of esophageal cancers. NedTijdschrTandheelkd 117(9):427–431Google Scholar
- 7.Ferlay J, Soerjomataram I, Ervik M, (2013) GLOBOCAN 2012 cancer incidence and mortality worldwide: IARC cancer base no. 11. International Agency for Research on Cancer. Accessed Jan 2016Google Scholar
- 9.Siewert JR, Stein HJ, Feith M, Bruecher BL, Bartels H, Fink U (2001) Histologic tumor type is an independent prognostic parameter in esophageal cancer: lessons from more than 1,000 consecutive resections at a single center in the Western world. Ann Surg 234(3):360–367 discussion 368-369PubMedPubMedCentralCrossRefGoogle Scholar
- 16.Bastid J, Regairaz A, Bonnefoy N, Dejou C, Giustiniani J, Laheurte C, Cochaud S, Laprevotte E, Funck-Brentano E, Hemon P, Gros L, Bec N, Larroque C, Alberici G, Bensussan A, Eliaou JF (2015) Inhibition of CD39 enzymatic function at the surface of tumor cells alleviates their immunosuppressive activity. Cancer immunology research 3(3):254–265. doi: 10.1158/2326-6066.CIR-14-0018 PubMedCrossRefGoogle Scholar
- 26.Gehring MP, Pereira TC, Zanin RF, Borges MC, Filho AB, Battastini AM, Bogo MR, Lenz G, Campos MM, Morrone FB (2012) P2X7 receptor activation leads to increased cell death in a radiosensitive human glioma cell line. Purinergic Signal 8(4):729–739. doi: 10.1007/s11302-012-9319-2 PubMedPubMedCentralCrossRefGoogle Scholar
- 31.Adinolfi E, Capece M, Franceschini A, Falzoni S, Giuliani AL, Rotondo A, Sarti AC, Bonora M, Syberg S, Corigliano D, Pinton P, Jorgensen NR, Abelli L, Emionite L, Raffaghello L, Pistoia V, DiVirgilio F (2015) Accelerated tumor progression in mice lacking the ATP receptor P2X7. Cancer Res 75(4):635–644. doi: 10.1158/0008-5472.CAN-14-1259 PubMedCrossRefGoogle Scholar
- 33.Vazquez-Cuevas FG, Martinez-Ramirez AS, Robles-Martinez L, Garay E, Garcia-Carranca A, Perez-Montiel D, Castaneda-Garcia C, Arellano RO (2014) Paracrine stimulation of P2X7 receptor by ATP activates a proliferative pathway in ovarian carcinoma cells. J Cell Biochem 115(11):1955–1966. doi: 10.1002/jcb.24867 PubMedGoogle Scholar
- 39.Michaud M, Martins I, Sukkurwala AQ, Adjemian S, Ma Y, Pellegatti P, Shen S, Kepp O, Scoazec M, Mignot G, Rello-Varona S, Tailler M, Menger L, Vacchelli E, Galluzzi L, Ghiringhelli F, di Virgilio F, Zitvogel L, Kroemer G (2011) Autophagy-dependent anticancer immune responses induced by chemotherapeutic agents in mice. Science 334(6062):1573–1577. doi: 10.1126/science.1208347 PubMedCrossRefGoogle Scholar
- 47.Adinolfi E, Cirillo M, Woltersdorf R, Falzoni S, Chiozzi P, Pellegatti P, Callegari MG, Sandona D, Markwardt F, Schmalzing G, Di Virgilio F (2010) Trophic activity of a naturally occurring truncated isoform of the P2X7 receptor. FASEB J 24(9):3393–3404. doi: 10.1096/fj. 09-153601 PubMedCrossRefGoogle Scholar
- 49.Gehring MP, Kipper F, Nicoletti NF, Sperotto ND, Zanin R, Tamajusuku AS, Flores DG, Meurer L, Roesler R, Filho AB, Lenz G, Campos MM, Morrone FB (2015) P2X7 receptor as predictor gene for glioma radiosensitivity and median survival. Int J Biochem Cell Biol 68:92–100. doi: 10.1016/j.biocel.2015.09.001 PubMedCrossRefGoogle Scholar
- 50.Ghiringhelli F, Apetoh L, Tesniere A, Aymeric L, Ma Y, Ortiz C, Vermaelen K, Panaretakis T, Mignot G, Ullrich E, Perfettini JL, Schlemmer F, Tasdemir E, Uhl M, Genin P, Civas A, Ryffel B, Kanellopoulos J, Tschopp J, Andre F, Lidereau R, McLaughlin NM, Haynes NM, Smyth MJ, Kroemer G, Zitvogel L (2009) Activation of the NLRP3 inflammasome in dendritic cells induces IL-1beta-dependent adaptive immunity against tumors. Nat Med 15(10):1170–1178. doi: 10.1038/nm.2028 PubMedCrossRefGoogle Scholar
- 66.Maaser K, Höpfner M, Kap H, Sutter AP, Barthel B, von Lampe B, Zeitz M, Scherübl H (2002) Extracellular nucleotides inhibit growth of human oesophageal cancer cells via P2Y(2)-receptors. Br Jn 86(4):636–644Google Scholar