Proapoptotic effect of endocannabinoids in prostate cancer cells

Orellana-Serradell,O.; Poblete,C. E.; Sanchez,C.; Castellón,E. A.; Gallegos,I.; Huidobro,C.; Llanos,M. N.; Contreras,H. R.

doi:10.3892/or.2015.3746

Proapoptotic effect of endocannabinoids in prostate cancer cells

Authors:
- O. Orellana-Serradell
- C. E. Poblete
- C. Sanchez
- E. A. Castellón
- I. Gallegos
- C. Huidobro
- M. N. Llanos
- H. R. Contreras
View Affiliations

Affiliations: Physiology and Biophysics Program, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago 8389100, Chile, Pathological Anatomy Service, Clinic Hospital of the University of Chile, Santiago 8389100, Chile, Urology Service, Clinic Hospital of the University of Chile, Santiago 8389100, Chile, Laboratory of Nutrition and Metabolic Regulation, INTA, University of Chile, Santiago 8389100, Chile
Published online on: January 21, 2015 https://doi.org/10.3892/or.2015.3746
Pages: 1599-1608
Copyright: © Orellana-Serradell et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

In the early stages, prostate cancer is androgen‑ dependent; therefore, medical castration has shown significant results during the initial stages of this pathology. Despite this early effect, advanced prostate cancer is resilient to such treatment. Recent evidence shows that derivatives of Cannabis sativa and its analogs may exert a protective effect against different types of oncologic pathologies. The purpose of the present study was to detect the presence of cannabinoid receptors (CB1 and CB2) on cancer cells with a prostatic origin and to evaluate the effect of the in vitro use of synthetic analogs. In order to do this, we used a commercial cell line and primary cultures derived from prostate cancer and benign prostatic hyperplasia. The presence of the CB1 and CB2 receptors was determined by immunohistochemistry where we showed a higher expression of these receptors in later stages of the disease (samples with a high Gleason score). Later, treatments were conducted using anandamide, 2-arachidonoyl glycerol and a synthetic analog of anandamide, methanandamide. Using the MTT assay, we proved that the treatments produced a cell growth inhibitory effect on all the different prostate cancer cultures. This effect was demonstrated to be dose-dependent. The use of a specific CB1 receptor blocker (SR141716) confirmed that this effect was produced primarily from the activation of the CB1 receptor. In order to understand the MTT assay results, we determined cell cycle distribution by flow cytometry, which showed no variation at the different cell cycle stages in all the cultures after treatment. Treatment with endocannabinoids resulted in an increase in the percentage of apoptotic cells as determined by Annexin V assays and caused an increase in the levels of activated caspase-3 and a reduction in the levels of Bcl-2 confirming that the reduction in cell viability noted in the MTT assay was caused by the activation of the apoptotic pathway. Finally, we observed that endocannabinoid treatment activated the Erk pathway and at the same time, produced a decrease in the activation levels of the Akt pathway. Based on these results, we suggest that endocannabinoids may be a beneficial option for the treatment of prostate cancer that has become nonresponsive to common therapies.

View Figures

View References

1	Jemal A, Bray F, Center MM, et al: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	Sark W: Epidemiology of prostate cancer and its precursors. Mod Pathol. 10:10382004.(Epub ahead of print). View Article : Google Scholar
3	Tefekli A and Tunc M: Future prospects in the diagnosis and management of localized prostate cancer. Sci World J. 2013:3472632013. View Article : Google Scholar
4	Hsing A and Chokkalingam A: Prostate cancer epidemiology. Front Biosci. 11:1388–1413. 2006. View Article : Google Scholar
5	Porkka K and Visakorpi T: Molecular mechanisms of prostate cancer. Eur Urol. 45:683–691. 2004. View Article : Google Scholar : PubMed/NCBI
6	Lee C, Jia Z, Rahmatpanah F, Zhang Q, Zi X, McClelland M and Mercola D: Role of the adjacent stroma cells in prostate cancer development and progression: synergy between TGF-β and IGF signaling. Biomed Res Int. 2014:5020932014.
7	Bifulco M, Malfitano A, Pisanti S and Laezza C: Endocannabinoids in endocrine and related tumours. Endocr Relat Cancer. 15:391–408. 2008. View Article : Google Scholar : PubMed/NCBI
8	Hermanson DJ and Marnett LJ: Cannabinoids, endocannabinoids, and cancer. Cancer Metastasis Rev. 30:599–612. 2011. View Article : Google Scholar : PubMed/NCBI
9	Díaz-Laviada I: The endocannabinoid system in prostate cancer. Nat Rev Urol. 8:553–561. 2011. View Article : Google Scholar : PubMed/NCBI
10	Devane WA, Dysarz FA, Johnson MR, et al: Determination and characterization of a cannabinoid receptor in rat brain. Mol Pharmacol. 34:605–613. 1998.
11	Van Dross R, Soliman E, Jha S, et al: Receptor-dependent and receptor-independent endocannabinoid signaling: A therapeutic target for regulation of cancer growth. Life Sci. 92:463–466. 2013. View Article : Google Scholar
12	Guindon J and Hohmann AG: The endocannabinoid system and cancer: therapeutic implication. Br J Pharmacol. 16:1447–1463. 2011. View Article : Google Scholar
13	Pisanti S and Bifulco M: Endocannabinoid system modulation in cancer biology and therapy. Pharmacol Res. 60:107–116. 2009. View Article : Google Scholar : PubMed/NCBI
14	Kirkham TC and Tucci SA: Endocannabinoids in appetite control and the treatment of obesity. CNS Neurol Disord Drug Targets. 5:272–292. 2006. View Article : Google Scholar : PubMed/NCBI
15	Bovolin P, Cottone E, Pomatto V, et al: Endocannabinoids are involved in male vertebrate reproduction: regulatory mechanisms at central and gonadal level. Front Endocrinol (Lausanne). 5:542014.
16	Jiang W, Zhang Y, Xiao L, Van Cleemput J, Ji SP, Bai G and Zhang X: Cannabinoids promote embryonic and adult hippocampus neurogenesis and produce anxiolytic- and antidepressant-like effects. J Clin Invest. 115:3104–3116. 2005. View Article : Google Scholar : PubMed/NCBI
17	Cridge BJ and Rosengren RJ: Critical appraisal of the potential use of cannabinoids in cancer management. Cancer Manag Res. 30:301–313. 2013.
18	Batista LA, Gobira PH, Viana TG, et al: Inhibition of endocannabinoid neuronal uptake and hydrolysis as strategies for developing anxiolytic drugs. Behav Pharmacol. 25:425–433. 2014.PubMed/NCBI
19	Bifulco M, Laezza C, Pisanti S and Gazzerro P: Cannabinoids and cancer: pros and cons of an antitumor strategy. Br J Pharmacol. 148:123–135. 2006. View Article : Google Scholar : PubMed/NCBI
20	Ramos JA and Bianco FJ: The role of cannabinoids in prostate cancer: Basic science perspective and potential clinical applications. Indian J Urol. 28:9–14. 2012. View Article : Google Scholar : PubMed/NCBI
21	Melck D, Rueda D, Galve-Roperh I, et al: Involvement of the cAMP/protein kinase A pathway and of mitogen-activated protein kinase in the anti-proliferative effects of anandamide in human breast cancer cells. FEBS Lett. 463:235–240. 1999. View Article : Google Scholar : PubMed/NCBI
22	Nithipatikom K, Endsley M, Isbell M, et al: 2-Arachidonoyl-glycerol: a novel inhibitor of androgen-independent prostate cancer cell invasion. Cancer Res. 15:8826–8830. 2004. View Article : Google Scholar
23	Portella G, Laezza C, Laccetti P, et al: Inhibitory effects of cannabinoid CB1 receptor stimulation on tumor growth and metastatic spreading: actions on signals involved in angiogenesis and metastasis. FASEB J. 17:1771–1773. 2003.PubMed/NCBI
24	Melck D, De Petrocellis L, Orlando P, et al: Suppression of nerve growth factor Trk receptor and prolactin receptors by endocannabinoids leads to inhibition of human breast and prostate cancer cell proliferation. Endocrinology. 141:118–126. 2000.
25	Czifra G, Varga A, Nyeste K, et al: Increased expressions of cannabinoid receptor-1 and transient receptor potential vanilloid-1 in human prostate carcinoma. J Cancer Res Clin Oncol. 135:507–514. 2009. View Article : Google Scholar
26	Mimeault M, Pommery N, Wattez N, et al: Anti-proliferative and apoptotic effects of anandamide in human prostatic cancer cell lines: implication of epidermal growth factor receptor down-regulation and ceramide production. Postate. 15:1–12. 2003. View Article : Google Scholar
27	Ruiz L, Miguel A and Díaz-Laviada I: Delta9-tetrahydro-cannabinol induces apoptosis in human prostate PC-3 cells via a receptor-independent mechanism. FEBS Lett. 458:400–404. 1999. View Article : Google Scholar : PubMed/NCBI
28	Sarfaraz S, Afaq F, Adhami VM and Mukhtar H: Cannabinoid receptor as a novel target for the treatment of prostate cancer. Cancer Res. 65:1635–1641. 2005. View Article : Google Scholar : PubMed/NCBI
29	Gómez del Pulgar T, Velasco G and Guzmán M: The CB1 cannabinoid receptor is coupled to the activation of protein kinase B/Akt. Biochem J. 347:369–373. 2000. View Article : Google Scholar : PubMed/NCBI
30	Gómez del Pulgar T, Velasco G, Sánchez C, et al: De novo-synthesized ceramide is involved in cannabinoid-induced apoptosis. Biochem J. 363:183–188. 2002. View Article : Google Scholar : PubMed/NCBI
31	Rueda D, Galve-Roperh I, Haro A and Guzmán M: The CB1 cannabinoid receptor is coupled to the activation of c-Jun N-terminal kinase. Mol Pharmacol. 58:814–820. 2000.PubMed/NCBI
32	Mendoza P, Sánchez C, Contreras HR, et al: Evaluation of MENT on primary cell cultures from benign prostatic hyperplasia and prostate carcinoma. Int J Androl. 32:607–615. 2009. View Article : Google Scholar
33	Chunga S, Hammarstenb P, Josefssonb A, et al: A high cannabinoid CB(1) receptor immunoreactivity is associated with disease severity and outcome in prostate cancer. Eur J Cancer. 45:174–182. 2009. View Article : Google Scholar
34	Brown L, Cascio M, Wahle K, et al: Cannabinoid receptor- dependent and -independent anti-proliferative effects of omega-3 ethanolamides in androgen receptor-positive and-negative prostate cancer cell lines. Carcinogenesis. 31:1584–1591. 2010. View Article : Google Scholar : PubMed/NCBI
35	Nithipatikom K, Isbell M, Endsleya M, et al: Anti-proliferative effect of a putative endocannabinoid, 2-arachidonylglyceryl ether in prostate carcinoma cells. Prostaglandins Other Lipid Mediat. 94:34–43. 2011. View Article : Google Scholar :
36	Olea-Herrero N, Vara D, Malagarie-Cazenave S and Díaz-Laviada I: Inhibition of human tumour prostate PC-3 cell growth by cannabinoids R(+)-Methanandamide and JWH-015: involvement of CB2. Br J Cancer. 101:940–950. 2009. View Article : Google Scholar : PubMed/NCBI
37	Agudelo M, Newton C, Widen R, et al: Cannabinoid receptor 2 (CB2) mediates immunoglobulin class switching from IgM to IgE in cultures of murine-purified B lymphocytes. J Neuroimmune Pharmacol. 3:35–42. 2008. View Article : Google Scholar : PubMed/NCBI
38	Kelman Z: PCNA: structure, function and interactions. Oncogene. 6:629–640. 1997. View Article : Google Scholar
39	Song G, Ouyang G and Bao S: The activation of Akt/PKB signaling pathway and cell survival. J Cell Mol Med. 1:59–71. 2005. View Article : Google Scholar
40	Greenhough A, Patsos H, Ann C, et al: The cannabinoid D9-tetrahydrocannabinol inhibits RAS-MAPK and PI3K-AKT survival signalling and induces BAD-mediated apoptosis in colorectal cancer cells. Int J Cancer. 121:2172–2180. 2007. View Article : Google Scholar : PubMed/NCBI
41	De Petrocellis L, Ligresti A, Moriello A, et al: Non-THC cannabinoids inhibit prostate carcinoma growth in vitro and in vivo: pro-apoptotic effects and underlying mechanisms. Br J Pharmacol. 168:79–102. 2013. View Article : Google Scholar :
42	Hsu SS, Huang CJ, Cheng HH, et al: Anandamide-induced Ca²⁺ elevation leading to p38 MAPK phosphorylation and subsequent cell death via apoptosis in human osteosarcoma cells. Toxicology. 28:21–29. 2007. View Article : Google Scholar