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Journal of Physiology and Biochemistry

, Volume 74, Issue 2, pp 261–272 | Cite as

Cannabinoid-induced cell death in endometrial cancer cells: involvement of TRPV1 receptors in apoptosis

  • B. M. FonsecaEmail author
  • G. Correia-da-Silva
  • N. A. Teixeira
Original Article

Abstract

Among a variety of phytocannabinoids, Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) are the most promising therapeutic compounds. Besides the well-known palliative effects in cancer patients, cannabinoids have been shown to inhibit in vitro growth of tumor cells. Likewise, the major endocannabinoids (eCBs), anandamide (AEA) and 2-arachidonoylglycerol (2-AG), induce tumor cell death. The purpose of the present study was to characterize cannabinoid elements and evaluate the effect of cannabinoids in endometrial cancer cell viability. The presence of cannabinoid receptors, transient receptor potential vanilloid 1 (TRPV1), and endocannabinoid-metabolizing enzymes were determined by qRT-PCR and Western blot. We also examined the effects and the underlying mechanisms induced by eCBs and phytocannabinoids in endometrial cancer cell viability. Besides TRPV1, both EC cell lines express all the constituents of the endocannabinoid system. We observed that at concentrations higher than 5 μM, eCBs and CBD induced a significant reduction in cell viability in both Ishikawa and Hec50co cells, whereas THC did not cause any effect. In Ishikawa cells, contrary to Hec50co, treatment with AEA and CBD resulted in an increase in the levels of activated caspase −3/−7, in cleaved PARP, and in reactive oxygen species generation, confirming that the reduction in cell viability observed in the MTT assay was caused by the activation of the apoptotic pathway. Finally, these effects were dependent on TRPV1 activation and intracellular calcium levels. These data indicate that cannabinoids modulate endometrial cancer cell death. Selective targeting of TPRV1 by AEA, CBD, or other stable analogues may be an attractive research area for the treatment of estrogen-dependent endometrial carcinoma. Our data further support the evaluation of CBD and CBD-rich extracts for the potential treatment of endometrial cancer, particularly, that has become non-responsive to common therapies.

Keywords

Endocannabinoids Phytocannabinoids Endometrial cancer Apoptosis 

Abbreviations

AEA

Anandamide

CB1

Cannabinoid receptor 1

CB2

Cannabinoid receptor 2

CBD

Cannabidiol

CPZ

Capsazepine

CPS

Capsaicin

2-AG

2-Arachidonoylglycerol

DMSO

Dimethylsulfoxide

E2

Estradiol

EC

Endometrial cancer

eCBs

Endocannabinoids

ER

Estrogen receptor

FAAH

Fatty acid amide hydrolase

iRTX

5’-Iodoresiniferatoxin

MAGL

Monoacylglycerol lipase

MTT

3-(4–5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NAPE

N-Arachidonoyl phosphatidylethanolamine

PI

Phosphatidylinositol

PR

Progesterone receptor

Notes

Funding information

The authors are grateful for the financial support under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), UID/MULTI/04378/2013-POCI/01/0145/FEDER/007728 from FCT/MEC through national funds and co-financed by FEDER - Portugal, under the Partnership Agreement PT2020, NORTE-01-0145-FEDER-000024.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Almada M, Amaral C, Diniz-da-Costa M, Correia-da-Silva G, Teixeira NA, Fonseca BM (2016) The endocannabinoid anandamide impairs in vitro decidualization of human cells. Reproduction 152(4):351–361.  https://doi.org/10.1530/REP-16-0364 CrossRefPubMedGoogle Scholar
  2. 2.
    Amant F, Moerman P, Neven P, Timmerman D, Van Limbergen E, Vergote I (2005) Endometrial cancer. Lancet 366:491–505.  https://doi.org/10.1016/S0140-6736(05)67063-8 CrossRefPubMedGoogle Scholar
  3. 3.
    Anzai Y, Gong YW, Holinka CF, Murphy LJ, Murphy LC, Kuramoto H, Gurpide E (1992) Effects of transforming growth-factors and regulation of their messenger-Rna levels in 2 human endometrial adenocarcinoma cell-lines. J Steroid Biochem Mol Biol 42:449–455.  https://doi.org/10.1016/0960-0760(92)90256-I CrossRefPubMedGoogle Scholar
  4. 4.
    Bartosch C, Gomes B, Jeronimo C, Lopes JM (2015) Endometrial carcinoma in Portugal: demographic, diagnostic and treatment changes in the last 5 decades. Int J Gynecol Pathol 34(2):159–168.  https://doi.org/10.1097/Pgp.0000000000000124 CrossRefPubMedGoogle Scholar
  5. 5.
    Bifulco M, Di Marzo V (2002) Targeting the endocannabinoid system in cancer therapy: a call for further research. Nat Med 8(6):547–550.  https://doi.org/10.1038/nm0602-547 CrossRefPubMedGoogle Scholar
  6. 6.
    Blazquez C, Carracedo A, Barrado L, Real PJ, Fernandez-Luna JL, Velasco G, Malumbres M, Guzman M (2006) Cannabinoid receptors as novel targets for the treatment of melanoma. Faseb J 20:2633-+.  https://doi.org/10.1096/fj.06-6638fje CrossRefPubMedGoogle Scholar
  7. 7.
    Brown I, Cascio MG, Rotondo D, Pertwee RG, Heys SD, Wahle KWJ (2013) Cannabinoids and omega-3/6 endocannabinoids as cell death and anticancer modulators. Prog Lipid Res 52(1):80–109.  https://doi.org/10.1016/j.plipres.2012.10.001 CrossRefPubMedGoogle Scholar
  8. 8.
    Colombo N, Preti E, Landoni F, Carinelli S, Colombo A, Marini C, Sessa C, Grp EGW (2013) Endometrial cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 24(suppl 6):33–38.  https://doi.org/10.1093/annonc/mdt353 CrossRefGoogle Scholar
  9. 9.
    Contassot E, Tenan M, Schnuriger V, Pelte MF, Dietrich PY (2004) Arachidonyl ethanolamide induces apoptosis of uterine cervix cancer cells via aberrantly expressed vanilloid receptor-1. Gynecol Oncol 93(1):182–188.  https://doi.org/10.1016/j.ygyno.2003.12.040 CrossRefPubMedGoogle Scholar
  10. 10.
    Contassot E, Wilmotte R, Tenan M, Belkouch MC, Schnuriger V, De Tribolet N, Bourkhardt K, Dietrich PY (2004) Arachidonylethanolamide induces apoptosis of human glioma cells through vanilloid receptor-1. J Neuropath Exp Neur 63(9):956–963.  https://doi.org/10.1093/jnen/63.9.956 CrossRefPubMedGoogle Scholar
  11. 11.
    Costa MA, Fonseca BM, Keating E, Teixeira NA, Correia-da-Silva G (2014) 2-arachidonoylglycerol effects in cytotrophoblasts: metabolic enzymes expression and apoptosis in BeWo cells. Reproduction 147(3):301–311.  https://doi.org/10.1530/REP-13-0563 CrossRefPubMedGoogle Scholar
  12. 12.
    Dai D, Wolf DM, Litman ES, White MJ, Leslie KK (2002) Progesterone inhibits human endometrial cancer cell growth and invasiveness: down-regulation of cellular adhesion molecules through progesterone B receptors. Cancer Res 62(3):881–886PubMedGoogle Scholar
  13. 13.
    Diaz-Laviada I (2011) The endocannabinoid system in prostate cancer. Nat Rev Urol 8(10):553–561.  https://doi.org/10.1038/nrurol.2011.130 CrossRefPubMedGoogle Scholar
  14. 14.
    Fonseca BM, Costa MA, Almada M, Correia-da-Silva G, Teixeira NA (2013) Endogenous cannabinoids revisited: a biochemistry perspective. Prostaglandins Other Lipid Mediat 102–103:13–30.  https://doi.org/10.1016/j.prostaglandins.2013.02.002 CrossRefPubMedGoogle Scholar
  15. 15.
    Galve-Roperh I, Sanchez C, Cortes ML, Gomez del Pulgar T, Izquierdo M, Guzman M (2000) Anti-tumoral action of cannabinoids: involvement of sustained ceramide accumulation and extracellular signal-regulated kinase activation. Nat Med 6(3):313–319.  https://doi.org/10.1038/73171 CrossRefPubMedGoogle Scholar
  16. 16.
    Guida M, Ligresti A, De Filippis D, D'Amico A, Petrosino S, Cipriano M, Bifulco G, Simonetti S, Orlando P, Insabato L, Nappi C, Di Spiezio Sardo A, Di Marzo V, Iuvone T (2010) The levels of the endocannabinoid receptor CB2 and its ligand 2-arachidonoylglycerol are elevated in endometrial carcinoma. Endocrinology 151(3):921–928.  https://doi.org/10.1210/en.2009-0883 CrossRefPubMedGoogle Scholar
  17. 17.
    Guindon J, Hohmann AG (2011) The endocannabinoid system and cancer: therapeutic implication. Br J Pharmacol 163(7):1447–1463.  https://doi.org/10.1111/j.1476-5381.2011.01327.x CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Holinka CF, Hata H, Kuramoto H, Gurpide E (1986) Effects of steroid hormones and antisteroids on alkaline phosphatase activity in human endometrial cancer cells (Ishikawa line). Cancer Res 46(6):2771–2774PubMedGoogle Scholar
  19. 19.
    Iannotti FA, Hill CL, Leo A, Alhusaini A, Soubrane C, Mazzarella E, Russo E, Whalley BJ, Di Marzo V, Stephens GJ (2014) Nonpsychotropic plant cannabinoids, cannabidivarin (CBDV) and cannabidiol (CBD), activate and desensitize transient receptor potential vanilloid 1 (TRPV1) channels in vitro: potential for the treatment of neuronal hyperexcitability. ACS Chem Neurosci 5(11):1131–1141.  https://doi.org/10.1021/cn5000524 CrossRefPubMedGoogle Scholar
  20. 20.
    Kassan S, Mechanick JI, Gurpide E (1989) Altered estrogen receptor system in estrogen-unresponsive human endometrial adenocarcinoma cells. J Steroid Biochem 33(3):327–333.  https://doi.org/10.1016/0022-4731(89)90320-8 CrossRefPubMedGoogle Scholar
  21. 21.
    Ligresti A, Moriello AS, Starowicz K, Matias I, Pisanti S, De Petrocellis L, Laezza C, Portella G, Bifulco M, Di Marzo V (2006) Antitumor activity of plant cannabinoids with emphasis on the effect of cannabidiol on human breast carcinoma. J Pharmacol Exp Ther 318(3):1375–1387.  https://doi.org/10.1124/jpet.106.105247 CrossRefPubMedGoogle Scholar
  22. 22.
    Lim MP, Devi LA, Rozenfeld R (2011) Cannabidiol causes activated hepatic stellate cell death through a mechanism of endoplasmic reticulum stress-induced apoptosis. Cell Death Dis 2:ARTN e170.  https://doi.org/10.1038/cddis.2011.52 CrossRefGoogle Scholar
  23. 23.
    Maccarrone M, Bab R, Biro T, Cabral GA, Dey SK, Di Marzo V, Konje JC, Kunos G, Mechoulam R, Pacher P, Sharkey KA, Zimmer A (2015) Endocannabinoid signaling at the periphery: 50 years after THC. Trends Pharmacol Sci 36(5):277–296.  https://doi.org/10.1016/j.tips.2015.02.008 CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Martinez N, Aban CE, Leguizamon GF, Damiano AE, Farina MG (2016) TPRV-1 expression in human preeclamptic placenta. Placenta 40:25–28.  https://doi.org/10.1016/j.placenta.2016.02.008 CrossRefPubMedGoogle Scholar
  25. 25.
    Newman TA, Bailey JL, Stocker LJ, Woo YL, Macklon NS, Cheong YC (2013) Expression of neuronal markers in the endometrium of women with and those without endometriosis. Hum Reprod 28(9):2502–2510.  https://doi.org/10.1093/humrep/det274 CrossRefPubMedGoogle Scholar
  26. 26.
    Nishida M (2002) The Ishikawa cells from birth to the present. Hum Cell 15(3):104–117.  https://doi.org/10.1111/j.1749-0774.2002.tb00105.x CrossRefPubMedGoogle Scholar
  27. 27.
    Pinton P, Giorgi C, Siviero R, Zecchini E, Rizzuto R (2008) Calcium and apoptosis: ER-mitochondria Ca2+ transfer in the control of apoptosis. Oncogene 27(50):6407–6418.  https://doi.org/10.1038/onc.2008.308 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Ross RA (2003) Anandamide and vanilloid TRPV1 receptors. Br J Pharmacol 140(5):790–801.  https://doi.org/10.1038/sj.bjp.0705467 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Shrivastava A, Kuzontkoski PM, Groopman JE, Prasad A (2011) Cannabidiol induces programmed cell death in breast cancer cells by coordinating the cross-talk between apoptosis and autophagy. Mol Cancer Ther 10(7):1161–1172.  https://doi.org/10.1158/1535-7163.MCT-10-1100 CrossRefPubMedGoogle Scholar
  30. 30.
    Soliman E, Van Dross R (2016) Anandamide-induced endoplasmic reticulum stress and apoptosis are mediated by oxidative stress in non-melanoma skin cancer: receptor-independent endocannabinoid signaling. Mol Carcinogen 55(11):1807–1821.  https://doi.org/10.1002/mc.22429 CrossRefGoogle Scholar
  31. 31.
    Taylor AH, Abbas MS, Habiba MA, Konje JC (2010) Histomorphometric evaluation of cannabinoid receptor and anandamide modulating enzyme expression in the human endometrium through the menstrual cycle. Histochem Cell Biol 133(5):557–565.  https://doi.org/10.1007/s00418-010-0695-9 CrossRefPubMedGoogle Scholar
  32. 32.
    Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65(2):87–108.  https://doi.org/10.3322/caac.21262 CrossRefPubMedGoogle Scholar

Copyright information

© University of Navarra 2018

Authors and Affiliations

  1. 1.UCIBIO, REQUIMTE, Departamento de Ciências Biológicas, Laboratório de Bioquímica, Faculdade de FarmáciaUniversidade do PortoPortoPortugal

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