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

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.

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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

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Funding

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.

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Correspondence to B. M. Fonseca.

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Fonseca, B.M., Correia-da-Silva, G. & Teixeira, N.A. Cannabinoid-induced cell death in endometrial cancer cells: involvement of TRPV1 receptors in apoptosis. J Physiol Biochem 74, 261–272 (2018). https://doi.org/10.1007/s13105-018-0611-7

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Keywords

  • Endocannabinoids
  • Phytocannabinoids
  • Endometrial cancer
  • Apoptosis