Abstract 8: Anandamide reduces cell migration, invasion, and induces apoptosis in an in vitro model of prostate cancer
Authors: Roberto Domenica , Klotz Laurence , Venkateswaran Vasundara 
Published: July 2017

Abstract
Introduction and Objective: Prostate cancer (PCa) is the most commonly diagnosed cancer in men and the second leading cause of cancer related death in Canada. A large body of evidence supports a possible role for cannabinoids in certain aspects of human health and disease, acting as palliative agents as well as inhibitors of cancer cell proliferation, migration, invasion, and the angiogenesis of tumours. Anandamide (AEA) is a highly potent endogenously produced compound that acts as an agonist of the cannabinoid receptors (CB1 and CB2) as well as the transient receptor potential vanilloid receptor 1. AEA is mainly metabolized by fatty acid amide hydrolase (FAAH) and overexpression of the enzyme in PCa tissues is correlated with greater invasivity, malignancy and increasing Gleason score. Majority of studies have focused on the effect of AEA treatment on cell proliferation, without much focus on prostate cancer biology. This study aims to determine the anti-cancer effect of AEA on PCa cells. We hypothesize that anandamide will reduce migration, invasion, and induce apoptosis in PCa cells.
Methodology: Human PCa cells (DU145, LNCaP, and PC3) were treated with AEA at concentrations ranging from 1μM to 40μM and growth of cells (using the MTS assay) was assessed at various times. Wound-healing assays were conducted to investigate the migratory potential of cells following 24hr exposure to treatment, and trans-well invasion assays were performed to explore the influence of AEA on cell invasion. Flow cytometry, using the FITC Annexin V dead cell apoptosis kit, was used to detect the proportion of apoptotic versus proliferating cells following anandamide treatment.
Results: There was a significant reduction in the proliferation of DU145 cells at concentrations of 20μM and 40μM AEA compared to controls (p=0.02, 0.005). LNCaP cells on the other hand required a higher concentration of AEA to achieve this effect (p=0.05). Cell migration and invasion studies on DU145 cells revealed a significant reduction in cell motility at 20μM and 40μM AEA (p=0.006, 0.0005) as well as a significant reduction in cell invasion at 20μM and 40μM ((p=0.0002, 0.0001). Flow cytometry results indicate a significant increase in the proportion of apoptotic cells compared to control (3 fold change) when treated with 20μM and 40μM in DU145 and LNCaP cells.
Conclusion: We have shown that AEA has greatest significant influence on cell migration, invasion, and apoptosis. Further analysis on the role of AEA in these pathways is underway exploring alterations in expression levels of key proteins implicated in cell migration and apoptosis.
Funding/ Conflicts of Interest: None
Note: This abstract was not presented at the meeting.
Citation Format: Domenica Roberto, Laurence Klotz, Vasundara Venkateswaran. Anandamide reduces cell migration, invasion, and induces apoptosis in an in vitro model of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 8. doi:10.1158/1538-7445.AM2017-8