Arachidonyl ethanolamide induces apoptosis of uterine cervix cancer cells via aberrantly expressed vanilloid receptor-1.
Source
Oncology Division, Laboratory of Tumor Immunology, University Hospital, Geneva, Switzerland.
Abstract
OBJECTIVES:
Delta(9)-Tetrahydrocannabinol, the active agent of Cannabis sativa, exhibits well-documented antitumor properties, but little is known about the possible effects mediated by endogenous cannabinoids on human tumors. In the present study, we analyzed the effect of arachidonyl ethanolamide (AEA) on cervical carcinoma (CxCa) cell lines.
METHODS:
To assess the sensitivity of CxCa cells to AEA, we selected three cell lines that were exposed to increasing doses of AEA with or without antagonists to receptors to AEA. DNA fragmentation and caspase-7 activity were used as apoptosis markers. The expression of receptors to AEA were analyzed in CxCa cell lines as well as CxCa biopsies.
RESULTS:
The major finding was that AEA induced apoptosis of CxCa cell lines via aberrantly expressed vanilloid receptor-1, whereas AEA binding to the classical CB1 and CB2 cannabinoid receptors mediated a protective effect. Furthermore, unexpectedly, a strong expression of the three forms of AEA receptors was observed in ex vivo CxCa biopsies.
CONCLUSIONS:
Overall, these data suggest that the specific targeting of VR1 by endogenous cannabinoids or synthetic molecules offers attractive opportunities for the development of novel potent anticancer drugs.
- PMID:
- 15047233
- [PubMed – indexed for MEDLINE]
Publication Types, MeSH Terms, Substances
Publication Types
MeSH Terms
- Apoptosis/drug effects*
- Arachidonic Acids/pharmacology*
- Cannabinoid Receptor Modulators/pharmacology*
- Cell Line, Tumor
- Endocannabinoids
- Female
- HeLa Cells
- Humans
- Polyunsaturated Alkamides
- Receptor, Cannabinoid, CB1/biosynthesis
- Receptor, Cannabinoid, CB1/physiology
- Receptor, Cannabinoid, CB2/biosynthesis
- Receptor, Cannabinoid, CB2/physiology
- Receptors, Drug/biosynthesis
- Receptors, Drug/physiology*
- TRPV Cation Channels
- Uterine Cervical Neoplasms/drug therapy*
- Uterine Cervical Neoplasms/metabolism
- Uterine Cervical Neoplasms/pathology*
Substances
LinkOut – more resources
Full Text Sources
Other Literature Sources
- COS Scholar Universe
- Histopathology and cytopathology of the uterine cervix – digital atlas – International Agency for Research on Cancer – Screening Group
- A practical manual on visual screening for cervical neoplasia – International Agency for Research on Cancer – Screening Group
Medical
Figures and tables from this article:
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Fig. 1.
AEA-induced apoptosis of CxCa cell lines. (A) The viability of healthy donor PBLs (black square), CC299 (black triangle), Caski (white circle) and Hela cells (black circle) was assessed in a MTT assay after 5 days of AEA exposure. AEA was added daily in culture medium because of its high instability. Vehicle consisted in ethanol and never exceeded 1% of the final culture medium volume. Results are expressed as mean ± SD of triplicates. (B) Flow cytometry analysis of DNA fragmentation of CC299 cell line exposed to 30 μM AEA for 0, 15, 24, 48 and 72 h. Similar DNA content patterns were obtained with Caski and HeLa cells (percentages of cells in sub-G0/G1 reported in C for each cell line). (C) Kinetics analysis of the sub-G0/G1 cell fraction (fragmented DNA) in CxCa cells treated with 30 μM AEA (black circle) or vehicle (white circle). Results are expressed as mean ± SD of duplicates. (D) Western blot analysis of caspase-7 activation in CC299 cells. Cleaved form of caspase-7 (20 kDa) was observed 48 h after addition of 30 μM AEA.
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Fig. 2.
CB1, CB2 and VR1 are expressed by CxCa cell lines. Expression of cannabinoid receptors was assessed by RT-PCR (A) and Western blot analysis (B). Expected protein size was 84 kDa for VR1, 52 kDa for CB1 and 38 kDa for CB2.
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Fig. 3.
VR1 is involved in AEA-induced death of cervical cancer cells, whereas CB1 and CB2 are protective. Viability of CC299, Caski and HeLa cell lines was assessed by MTT assay after 3 days (black histogram) or 5 days (gray histogram) of AEA exposure. Antagonists to CB1 (SR1, 0.2 μM), CB2 (SR2, 0.2 μM) and VR1 (capsazepine, CZ, 0.5 μM) were added 15 min before addition of 30 μM AEA. Results are expressed as mean ± SD of triplicates.
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Fig. 4.
Expression of receptors to AEA in cervix malignancies. RT-PCR analysis using specific primers for VR1, CB1 and CB2 was performed on stage Ib–II CxCa biopsies.
- Corresponding author. Laboratory of Tumor Immunology, Oncology Division, University Hospital, Rue Micheli-du-Crest 24, 1211 Geneva 14, Switzerland. Fax: +41-22-372-98-58.
Copyright © 2004 Elsevier Inc. All rights reserved.
