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Involvement of the CB2 cannabinoid receptor in cell growth inhibition and G0/G1 cell cycle arrest via the cannabinoid agonist WIN 55,212-2 in renal cell carcinoma.
BMC Cancer 2018; 18(1):583BC

Abstract

BACKGROUND

The anti-tumor properties of cannabinoids have been investigated in many in vitro and in vivo studies. Many of these anti-tumor effects are mediated via cannabinoid receptor types 1 and 2 (CB1 and CB2), comprising the endocannabinoid system (ECS). In this study, we investigated the ECS based on CB 1 and CB 2 receptor gene and protein expression in renal cell carcinoma (RCC) cell lines. In view of their further use for potential treatments, we thus investigated the roles of CB1 and CB2 receptors in the anti-proliferative action and signal transduction triggered by synthetic cannabinoid agonists [such as JWH-133 and WIN 55,212-2 (WIN-55)] in RCC cell lines.

METHODS

Human RCC cell lines were used for this study. The CB 1 and CB 2 gene expression levels were analyzed using real-time PCR. Flow cytometric, immunocytochemical and western blot analyses were performed to confirm CB1 and CB2 receptor protein expression. The anti-proliferative effects of synthetic cannabinoids were investigated on cell viability assay. The CB1 and CB2 receptors were blocked pharmacologically with the antagonists SR141716A and AM-630, respectively, to investigate the effects of the agonists JWH-133 and WIN-55. Cell cycle, apoptosis and LDH-based cytotoxicity were analyzed on cannabinoid-treated RCC cells.

RESULTS

The CB1 and CB2 genes expression was shown by real-time PCR and flow cytometric and western blot analysis indicating a higher level of CB2 receptor as compared to CB1 in RCC cells. Immunocytochemical staining also confirmed the expression of the CB1 and CB2 proteins. We also found that the synthetic cannabinoid agonist WIN-55 exerted anti-proliferative and cytotoxic effects by inhibiting the growth of RCC cell lines, while the CB2 agonist JWH-133 did not. Pharmacologically blocking the CB1 and CB2 receptors with their respective antagonists SR141716A and AM-630, followed by the WIN-55 treatment of RCC cells allowed uncovering the involvement of CB2, which led to an arrest in the G0/G1 phase of the cell cycle and apoptosis.

CONCLUSIONS

This study elucidated the involvement of CB2 in the in vitro inhibition of RCC cells, and future applications of CB2 agonists in the prevention and management of RCC are discussed.

Authors+Show Affiliations

Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland. imrankhanbioinfo@gmail.com. Department of Otolaryngology - Head & Neck Surgery, Western University, London, ON, N6A 3K7, Canada. imrankhanbioinfo@gmail.com.Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland. Faculty of Biology, Warsaw University, ul. Miecznikowa 1, 02-096, Warsaw, Poland.Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.Department of Physiological Sciences, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159, 02-776, Warsaw, Poland.Department of Physiological Sciences, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159, 02-776, Warsaw, Poland.Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland. Warsaw Medical University, Żwirki i Wigury 61, 02-091, Warsaw, Poland.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29792186

Citation

Khan, Mohammed I., et al. "Involvement of the CB2 Cannabinoid Receptor in Cell Growth Inhibition and G0/G1 Cell Cycle Arrest Via the Cannabinoid Agonist WIN 55,212-2 in Renal Cell Carcinoma." BMC Cancer, vol. 18, no. 1, 2018, p. 583.
Khan MI, Sobocińska AA, Brodaczewska KK, et al. Involvement of the CB2 cannabinoid receptor in cell growth inhibition and G0/G1 cell cycle arrest via the cannabinoid agonist WIN 55,212-2 in renal cell carcinoma. BMC Cancer. 2018;18(1):583.
Khan, M. I., Sobocińska, A. A., Brodaczewska, K. K., Zielniok, K., Gajewska, M., Kieda, C., ... Szczylik, C. (2018). Involvement of the CB2 cannabinoid receptor in cell growth inhibition and G0/G1 cell cycle arrest via the cannabinoid agonist WIN 55,212-2 in renal cell carcinoma. BMC Cancer, 18(1), p. 583. doi:10.1186/s12885-018-4496-1.
Khan MI, et al. Involvement of the CB2 Cannabinoid Receptor in Cell Growth Inhibition and G0/G1 Cell Cycle Arrest Via the Cannabinoid Agonist WIN 55,212-2 in Renal Cell Carcinoma. BMC Cancer. 2018 May 23;18(1):583. PubMed PMID: 29792186.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Involvement of the CB2 cannabinoid receptor in cell growth inhibition and G0/G1 cell cycle arrest via the cannabinoid agonist WIN 55,212-2 in renal cell carcinoma. AU - Khan,Mohammed I, AU - Sobocińska,Anna A, AU - Brodaczewska,Klaudia K, AU - Zielniok,Katarzyna, AU - Gajewska,Malgorzata, AU - Kieda,Claudine, AU - Czarnecka,Anna M, AU - Szczylik,Cezary, Y1 - 2018/05/23/ PY - 2017/01/31/received PY - 2018/05/09/accepted PY - 2018/5/25/entrez PY - 2018/5/25/pubmed PY - 2019/1/15/medline KW - CB1 and CB2 receptors KW - Endocannabinoid system (ECS) KW - JWH-133 KW - Renal cell carcinoma KW - WIN 55,212–2 mesylate SP - 583 EP - 583 JF - BMC cancer JO - BMC Cancer VL - 18 IS - 1 N2 - BACKGROUND: The anti-tumor properties of cannabinoids have been investigated in many in vitro and in vivo studies. Many of these anti-tumor effects are mediated via cannabinoid receptor types 1 and 2 (CB1 and CB2), comprising the endocannabinoid system (ECS). In this study, we investigated the ECS based on CB 1 and CB 2 receptor gene and protein expression in renal cell carcinoma (RCC) cell lines. In view of their further use for potential treatments, we thus investigated the roles of CB1 and CB2 receptors in the anti-proliferative action and signal transduction triggered by synthetic cannabinoid agonists [such as JWH-133 and WIN 55,212-2 (WIN-55)] in RCC cell lines. METHODS: Human RCC cell lines were used for this study. The CB 1 and CB 2 gene expression levels were analyzed using real-time PCR. Flow cytometric, immunocytochemical and western blot analyses were performed to confirm CB1 and CB2 receptor protein expression. The anti-proliferative effects of synthetic cannabinoids were investigated on cell viability assay. The CB1 and CB2 receptors were blocked pharmacologically with the antagonists SR141716A and AM-630, respectively, to investigate the effects of the agonists JWH-133 and WIN-55. Cell cycle, apoptosis and LDH-based cytotoxicity were analyzed on cannabinoid-treated RCC cells. RESULTS: The CB1 and CB2 genes expression was shown by real-time PCR and flow cytometric and western blot analysis indicating a higher level of CB2 receptor as compared to CB1 in RCC cells. Immunocytochemical staining also confirmed the expression of the CB1 and CB2 proteins. We also found that the synthetic cannabinoid agonist WIN-55 exerted anti-proliferative and cytotoxic effects by inhibiting the growth of RCC cell lines, while the CB2 agonist JWH-133 did not. Pharmacologically blocking the CB1 and CB2 receptors with their respective antagonists SR141716A and AM-630, followed by the WIN-55 treatment of RCC cells allowed uncovering the involvement of CB2, which led to an arrest in the G0/G1 phase of the cell cycle and apoptosis. CONCLUSIONS: This study elucidated the involvement of CB2 in the in vitro inhibition of RCC cells, and future applications of CB2 agonists in the prevention and management of RCC are discussed. SN - 1471-2407 UR - https://www.unboundmedicine.com/medline/citation/29792186/Involvement_of_the_CB2_cannabinoid_receptor_in_cell_growth_inhibition_and_G0/G1_cell_cycle_arrest_via_the_cannabinoid_agonist_WIN_55212_2_in_renal_cell_carcinoma_ L2 - https://bmccancer.biomedcentral.com/articles/10.1186/s12885-018-4496-1 DB - PRIME DP - Unbound Medicine ER -