Abstract
Ovarian cancer is the deadliest gynecologic cancer due to lack of early effective diagnosis and development of resistance to platinum-based chemotherapy. Several studies reported that adenosine concentrations are higher in tumor microenvironment than in non-tumor tissue. This finding inspired us to study the role of adenosine in ovarian cancer cells and to investigate if adenosine pathways offer new treatment options urgently needed to prevent or overcome chemoresistance. The ovarian cancer cell lines HEY, A2780, and its cisplatin-resistant subline A2780CisR were used in this study. Expression and functional activity of adenosine receptors were investigated by RT-PCR, Western blotting, and cAMP assay. A1 and A2B adenosine receptors were expressed and functionally active in all three cell lines. Adenosine showed moderate cytotoxicity (MTT-IC50 values were between 700 and 900 μM) and induced apoptosis in a concentration-dependent manner by increasing levels of sub-G1 and cleaved PARP. Apoptosis was diminished by QVD-OPh, confirming caspase-dependent induction of apoptosis. Forty-eight hours pre-incubation of adenosine prior to cisplatin significantly enhanced cisplatin-induced cytotoxicity in a synergistic manner and increased apoptosis. SLV320 or PSB603, selective A1 and A2B antagonists, was not able to inhibit adenosine-induced increase in cisplatin cytotoxicity or apoptosis whereas dipyridamole, a nucleoside transporter inhibitor, completely abrogated both effects. Mechanistically, adenosine increased pAMPK and reduced pS6K which was prevented by dipyridamole. In conclusion, application of adenosine prior to cisplatin could be a new therapeutic option to increase the potency of cisplatin in a synergistic manner and thus overcome platinum resistance in ovarian cancer.
Similar content being viewed by others
References
Coward JI, Middleton K, Murphy F (2016) New perspectives on targeted therapy in ovarian cancer. Int J Womens Health 7:189–203
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM (2010) Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 127(12):2893–2917
Ledermann JA, Raja FA, Fotopoulou C, Gonzalez-Martin A, Colombo N, Sessa C (2013;24 Suppl 6) Newly diagnosed and relapsed epithelial ovarian carcinoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol:vi24–vi32
Foley OW, Rauh-Hain JA, Carmen MG (2013) Recurrent epithelial ovarian cancer: an update on treatment. Oncology (Williston Park) 27(4):288–294 298
Mantia-Smaldone GM, Edwards RP, Vlad AM (2011) Targeted treatment of recurrent platinum-resistant ovarian cancer: current and emerging therapies. Cancer Manag Res 3:25–38
Siddik ZH (2003) Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene 22:7265–7279
Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, Castedo M, Kroemer G (2012) Molecular mechanisms of cisplatin resistance. Oncogene 31(15):1869–1883
Hijaz M, Chhina J, Mert I, Taylor M, Dar S, Al-Wahab Z et al (2016) Preclinical evaluation of olaparib and metformin combination in BRCA1 wild type ovarian cancer. Gynecol Oncol 142(2):323–331
Kim G, Ison G, McKee AE, Zhang H, Tang S, Gwise T et al (2015) FDA approval summary: olaparib monotherapy in patients with deleterious germline BRCA-mutated advanced ovarian cancer treated with three or more lines of chemotherapy. Clin Cancer Res 21(19):4257–4261
Di Virgilio F, Adinolfi E (2017) Extracellular purines, purinergic receptors and tumor growth. Oncogene 36(3):293–303
Kumar V (2013) Adenosine as an endogenous immunoregulator in cancer pathogenesis: where to go? Purinergic Signal 9(2):145–165
Blay J, White TD, Hoskin DW (1997) The extracellular fluid of solid carcinomas contains immunosuppressive concentrations of adenosine. Cancer Res 57(13):2602–2605
US Food & Drug Administration (USFDA). Adenosine. https://www.accessdata.fda.gov. Accessed 27 July 2017
Gessi S, Merighi S, Sacchetto V, Simioni C, Borea PA (2011) Adenosine receptors and cancer. Biochim Biophys Acta 1808(5):1400–1412
Antonioli L, Blandizzi C, Pacher P, Hasko G (2013) Immunity, inflammation and cancer: a leading role for adenosine. Nat Rev Cancer 13(12):842–857
Dasgupta B, Chhipa RR (2016) Evolving lessons on the complex role of AMPK in normal physiology and cancer. Trends Pharmacol Sci 37(3):192–206
Ma XM, Blenis J (2009) Molecular mechanisms of mTOR-mediated translational control. Nat Rev Mol Cell Biol 10(5):307–318
Watanabe R, Wei L, Huang J (2011) mTOR signaling, function, novel inhibitors, and therapeutic targets. J Nucl Med 52(4):497–500
Liu H, Scholz C, Zang C, Schefe JH, Habbel P, Regierer AC, Schulz CO, Possinger K, Eucker J (2012) Metformin and the mTOR inhibitor everolimus (RAD001) sensitize breast cancer cells to the cytotoxic effect of chemotherapeutic drugs in vitro. Anticancer Res 32(5):1627–1637
William WN, Kim JS, Liu DD, Solis L, Behrens C, Lee JJ, Lippman SM, Kim ES, Hong WK, Wistuba II, Lee HY (2012) The impact of phosphorylated AMP-activated protein kinase expression on lung cancer survival. Ann Oncol 23(1):78–85
Zadra G, Batista JL, Loda M (2015) Dissecting the dual role of AMPK in cancer: from experimental to human studies. Mol Cancer Res 13(7):1059–1072
Liang J, Mills GB (2013) AMPK: a contextual oncogene or tumor suppressor? Cancer Res 73(10):2929–2935
Koumenis C, Hammond E, Giaccia A (2014) Tumor microenvironment and cellular stress: signaling, metabolism, imaging, and therapeutic targets. Preface. Adv Exp Med Biol 772:v–viii
Kim H, Hwang J, Yun H et al (2008) Inhibition of AMPK-activated protein kinase sensitizes cancer cells to cisplatin-induced apoptosis via hyper-induction of p53. J Biol Chem 283:3731–3742
Engelke LH, Hamacher A, Proksch P, Kassack MU (2016) Ellagic acid and resveratrol prevent the development of cisplatin resistance in the epithelial ovarian cancer cell line A2780. J Cancer 7(4):353–363
Stenzel K, Hamacher A, Hansen FK, Gertzen CGW, Senger J, Marquardt V, Marek L, Marek M, Romier C, Remke M, Jung M, Gohlke H, Kassack MU, Kurz T (2017) Alkoxyurean-based histone deacetylase inhibitors increase cisplatin potency in chemoresistant cancer cell lines. J Med Chem 60(13):5334–5348
Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden T (2012) Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13:134
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675
Kalk P, Eggert B, Relle K, Godes M, Heiden S, Sharkovska Y, Fischer Y, Ziegler D, Bielenberg GW, Hocher B (2007) The adenosine A1 receptor antagonist SLV320 reduces myocardial fibrosis in rats with 5/6 nephrectomy without affecting blood pressure. Br J Pharmacol 151(7):1025–1032
Chou TC (2006) Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 58(3):621–681
Gwinn DM, Shackelford DB, Egan DF, Mihaylova MM, Mery A, Vasquez DS, Turk BE, Shaw RJ (2008) AMPK phosphorylation of raptor mediates a metabolic checkpoint. Mol Cell 30(2):214–226
Shirali S, Aghaei M, Shabani M, Fathi M, Sohrabi M, Moeinifard M (2013) Adenosine induces cell cycle arrest and apoptosis via cyclinD1/Cdk4 and Bcl-2/Bax pathways in human ovarian cancer cell line OVCAR-3. Tumour Biol 34(2):1085–1095
Hajiahmadi S, Panjehpour M, Aghaei M, Mousavi S (2015a) Molecular expression of adenosine receptors in OVCAR-3, Caov-4 and SKOV-3 human ovarian cancer cell lines. Res Pharm Sci 10(1):43–51
Hajiahmadi S, Panjehpour M, Aghaei M, Shabani M (2015b) Activation of A2b adenosine receptor regulates ovarian cancer cell growth: involvement of Bax/Bcl-2 and caspase-3. Biochem Cell Biol 93(4):321–329
Mello Pde A, Filippi-Chiela EC, Nascimento J, Beckenkamp A, Santana DB, Kipper F et al (2014) Adenosine uptake is the major effector of extracellular ATP toxicity in human cervical cancer cells. Mol Biol Cell 25(19):2905–2918
Mello Pde A, Coutinho-Silva R, Savio LEB (2017) Multifaceted effects of extracellular adenosine triphosphate and adenosine in the tumor–host interaction and therapeutic perspectives. Frontier Immunol 8 article 1526
Wei Q, Costanzi S, Balasubramanian R, Gao ZG, Jacobson KA (2013) A2B adenosine receptor blockade inhibits growth of prostate cancer cells. Purinergic Signal 9(2):271–280
Mirza A, Basso A, Black S, Malkowski M, Kwee L, Pachter JA et al (2005) RNA interference targeting of A1 receptor-overexpressing breast carcinoma cells leads to diminished rates of cell proliferation and induction of apoptosis. Cancer Biol Ther 4(12):1355–1360
Ahmad A, Ahmad S, Glover L, Miller SM, Shannon JM, Guo X, Franklin WA, Bridges JP, Schaack JB, Colgan SP, White CW (2009) Adenosine A2A receptor is a unique angiogenic target of HIF-2alpha in pulmonary endothelial cells. Proc Natl Acad Sci 106(26):10684–10689
Ohta A, Gorelik E, Prasad SJ, Ronchese F, Lukashev D, Wong MK et al (2006) A2A adenosine receptor protects tumors from antitumor T cells. Proc Natl Acad Sci 103(35):13132–13137
Montalban Del Barrio I, Penski C, Schlahsa L, Stein RG, Diessner J, Wockel A et al (2016) Adenosine-generating ovarian cancer cells attract myeloid cells which differentiate into adenosine-generating tumor associated macrophages—a self-amplifying, CD39- and CD73-dependent mechanism for tumor immune escape. J Immunother Cancer 4:49
Domcke S, Sinha R, Levine DA, Sander C, Schultz N (2013) Evaluating cell lines as tumour models by comparison of genomic profiles. Nat Commun 4:2126
Gershenson DM (2016) Low-grade serous carcinoma of the ovary or peritoneum. Ann Oncol 27(suppl 1):i45–i49
Fernández ML, DiMattia GE, Dawson A, Bamford S, Anderson S, Hennessy BT, Anglesio MS, Shepherd TG, Salamanca C, Hoenisch J, Tinker A, Huntsman DG, Carey MS (2016) Differences in MEK inhibitor efficacy in molecularly characterized low-grade serous ovarian cancer cell lines. Am J Cancer Res 6(10):2235–2251
Virtanen SS, Kukkonen-Macchi A, Vainio M, Elima K, Harkonen PL, Jalkanen S, Yegutkin GG (2014) Adenosine inhibits tumor cell invasion via receptor-independent mechanisms. Mol Cancer Res 12(12):1863–1874
Yang D, Yaguchi T, Nakano T, Nishizaki T (2011) Adenosine activates AMPK to phosphorylate Bcl-XL responsible for mitochondrial damage and DIABLO release in HuH-7 cells. Cell Physiol Biochem 27(1):71–78
Wu LF, Li GP, Feng JL, Pu ZJ (2006) Molecular mechanisms of adenosine-induced apoptosis in human HepG2 cells. Acta Pharmacol Sin 27(4):477–484
Saitoh M, Nagai K, Nakagawa K, Yamamura T, Yamamoto S, Nishizaki T (2004) Adenosine induces apoptosis in the human gastric cancer cells via an intrinsic pathway relevant to activation of AMP-activated protein kinase. Biochem Pharmacol 67(10):2005–2011
Tsuchiya A, Nishizaki T (2015) Anticancer effect of adenosine on gastric cancer via diverse signaling pathways. World J Gastroenterol 21(39):10931–10935
Alers S, Loffler AS, Wesselborg S, Stork B (2012) Role of AMPK-mTOR-Ulk1/2 in the regulation of autophagy: cross talk, shortcuts, and feedbacks. Mol Cell Biol 32(1):2–11
Mihaylova MM, Shaw RJ (2011) The AMPK signaling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol 13(9):1016–1023
Chagoya de Sánchez V, Brunner A, Pina E (1972) In vivo modification of the energy charge in the liver cell. Biochem Biophys Res Commun 46(3):1441–1445
Oakhill JS, Steel R, Chen ZP, Scott JW, Ling N, Tam S, Kemp BE (2011) AMPK is a direct adenylate charge-regulated protein kinase. Science 332(6036):1433–1435
Tandon P, Gallo CA, Khatri S, Barger JF, Yepiskoposyan H, Plas DR (2011) Requirement for ribosomal protein S6 kinase 1 to mediate glycolysis and apoptosis resistance induced by Pten deficiency. Proc Natl Acad Sci U S A 108(6):2361–2365
Funding
This study was supported by the Bundesministerium für Forschung (BMBF, Germany, Grant: BMBF-16GW0108). We acknowledge funding by the DFG for the Thermofisher Arrayscan XTI (Grant: INST 208/690-1 FUGG).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflicts of interest
All the authors declare that they have no conflict of interest.
Ethical approval
This article does not contain any studies with participants or animals performed by any of the authors.
Electronic supplementary material
ESM 1
(DOCX 35 kb)
Rights and permissions
About this article
Cite this article
Sureechatchaiyan, P., Hamacher, A., Brockmann, N. et al. Adenosine enhances cisplatin sensitivity in human ovarian cancer cells. Purinergic Signalling 14, 395–408 (2018). https://doi.org/10.1007/s11302-018-9622-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11302-018-9622-7