¹Division of Experimental Medicine, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

Correspondence: Drs JE Groopman or RK Ganju, Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine Building, 4 Blackfan Circle, Boston, MA 02115, USA. E-mail:jgroopma@bidmc.harvard.edu or rganju@bidmc.harvard.edu

²JEG and RKG share the senior and corresponding authorship.

Received 21 January 2007; Revised 29 May 2007; Accepted 1 June 2007; Published online 9 July 2007.

Abstract

⁹-Tetrahydrocannabinol (THC) is the primary cannabinoid of marijuana and has been shown to either potentiate or inhibit tumor growth, depending on the type of cancer and its pathogenesis. Little is known about the activity of cannabinoids like THC on epidermal growth factor receptor-overexpressing lung cancers, which are often highly aggressive and resistant to chemotherapy. In this study, we characterized the effects of THC on the EGF-induced growth and metastasis of human non-small cell lung cancer using the cell lines A549 and SW-1573 as in vitro models. We found that these cells express the cannabinoid receptors CB₁ and CB₂, known targets for THC action, and that THC inhibited EGF-induced growth, chemotaxis and chemoinvasion. Moreover, signaling studies indicated that THC may act by inhibiting the EGF-induced phosphorylation of ERK1/2, JNK1/2 and AKT. THC also induced the phosphorylation of focal adhesion kinase at tyrosine 397. Additionally, in in vivo studies in severe combined immunodeficient mice, there was significant inhibition of the subcutaneous tumor growth and lung metastasis of A549 cells in THC-treated animals as compared to vehicle-treated controls. Tumor samples from THC-treated animals revealed antiproliferative and antiangiogenic effects of THC. Our study suggests that cannabinoids like THC should be explored as novel therapeutic molecules in controlling the growth and metastasis of certain lung cancers.

Experimental Cell Research  Volume 312, Issue 4, 15 February 2006, Pages 363-373

Abstract Full Text + Links PDF (383

doi:10.1016/j.yexcr.2005.10.024       
Copyright © 2005 Elsevier Inc. All rights reserved.

Research Article

Anandamide inhibits adhesion and migration of breast cancer cells

Claudia Grimaldi^a, 1, Simona Pisanti^a, 1, Chiara Laezza^b, Anna Maria Malfitano^a, Antonietta Santoro^a, Mario Vitale^c, Maria Gabriella Caruso^d, Maria Notarnicola^d, Irma Iacuzzo^a, b, Giuseppe Portella^b, Vincenzo Di Marzo^e, ,  and Maurizio Bifulco^a, ,  
aDipartimento di Scienze Farmaceutiche, Endocannabinoid Research Group, Università degli Studi di Salerno, Fisciano (Sa), Italy
^bIstituto di Endocrinologia e Oncologia Sperimentale del CNR e Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università di Napoli Federico II, Italy
^cDipartimento di Endocrinologia ed Oncologia Molecolare e Clinica, Università di Napoli Federico II, Italy
^dLaboratorio di Biochimica, IRCCS “S. de Bellis”, Castellana Grotte (Bari), Italy
^eIstituto di Chimica Biomolecolare, C.N.R., Pozzuoli (NA), Italy 
Received 6 May 2005;  revised 24 October 2005;  accepted 28 October 2005.  Available online 15 December 2005. 

Abstract

The endocannabinoid system regulates cell proliferation in human breast cancer cells. We reasoned that stimulation of cannabinoid CB₁ receptors could induce a non-invasive phenotype in breast mtastatic cells. In a model of metastatic spreading in vivo, the metabolically stable anandamide analogue, 2-methyl-2′-F-anandamide (Met-F-AEA), significantly reduced the number and dimension of metastatic nodes, this effect being antagonized by the selective CB₁ antagonist SR141716A. In MDA-MB-231 cells, a highly invasive human breast cancer cell line, and in TSA-E1 cells, a murine breast cancer cell line, Met-F-AEA inhibited adhesion and migration on type IV collagen in vitro without modifying integrin expression: both these effects were antagonized by SR141716A. In order to understand the molecular mechanism involved in these processes, we analyzed the phosphorylation of FAK and Src, two tyrosine kinases involved in migration and adhesion. In Met-F-AEA-treated cells, we observed a decreased tyrosine phosphorylation of both FAK and Src, this effect being attenuated by SR141716A. We propose that CB₁ receptor agonists inhibit tumor cell invasion and metastasis by modulating FAK phosphorylation, and that CB₁receptor activation might represent a novel therapeutic strategy to slow down the growth of breast carcinoma and to inhibit its metastatic diffusion in vivo.

Keywords: Endocannabinoid; Cancer; Metastasis

Corresponding authors. M. Bifulco is to be contacted at Dipartimento di Scienze Farmaceutiche, Università degli Studi di Salerno, Fisciano (Sa), Italy. V. Di Marzo, Istituto di Chimica Biomolecolare, C.N.R., Pozzuoli (NA), Italy.
¹ These authors contributed equally to the work.

Anandamide is an endogenous inhibitor for the migration of tumor cells and T lymphocytes Journal Cancer Immunology, Immunotherapy Publisher Springer Berlin / Heidelberg ISSN 0340-7004 (Print) 1432-0851 (Online) Issue Volume 53, Number 8 / August, 2004 Category Original Article DOI 10.1007/s00262-004-0509-9 Pages 723-728 Subject Collection Biomedical and Life Sciences SpringerLink Date Thursday, March 18, 2004

Jan Joseph1, Bernd Niggemann1, Kurt S. Zaenker1 and Frank Entschladen1  (1)  Institute of Immunology, Witten/Herdecke University, Stockumer Str. 10, 58448 Witten, Germany Received: 26 September 2003  Accepted: 27 January 2004  Published online: 18 March 2004 Abstract  Cell migration is of paramount importance in physiological processes such as immune surveillance, but also in the pathological processes of tumor cell migration and metastasis development. The factors that regulate this tumor cell migration, most prominently neurotransmitters, have thus been the focus of intense investigation. While the majority of neurotransmitters have a stimulatory effect on cell migration, we herein report the inhibitory effect of the endogenous substance anandamide on both tumor cell and lymphocyte migration. Using a collagen-based three-dimensional migration assay and time-lapse videomicroscopy, we have observed that the anandamide-mediated signals for CD8+ T lymphocytes and SW 480 colon carcinoma cells are each mediated by distinct cannabinoid receptors (CB-Rs). Using the specific agonist docosatetraenoylethanolamide (DEA), we have observed that the norepinephrine-induced migration of colon carcinoma cells is inhibited by the CB1-R. The SDF-1–induced migration of CD8+T lymphocytes was, however, inhibited via the CB2-R, as shown by using the specific agonist JWH 133. Therefore, specific inhibition of tumor cell migration via CB1-R engagement might be a selective tool to prevent metastasis formation without depreciatory effects on the immune system of cancer patients. Keywords  Anandamide - Cannabinoid receptors - Cell migration - T lymphocytes - Tumor cells The FASEB Journal Express Article doi:10.1096/fj.02-1129fje  Published online July 3, 2003 Inhibitory effects of cannabinoid CB1 receptor stimulation on tumor growth and metastatic spreading: actions on signals involved in angiogenesis and metastasis Giuseppe Portella, Chiara Laezza, Paolo Laccetti, Luciano De Petrocellis, Vincenzo Di Marzo, and Maurizio Bifulco E-mail contact: vdimarzo@icmib.na.cnr.it, maubiful@unina.it Stimulation of cannabinoid CB1 receptors by 2-methyl-arachidonyl-2′-fluoro-ethylamide (Met-F-AEA) inhibits the growth of a rat thyroid cancer cell-derived tumor in athymic mice by inhibiting the activity of the oncogene product p21ras. Here we report that Met-F-AEA also blocks the growth of tumors previously induced in nude mice by the s.c. injection of the same rat thyroid carcinoma cells. Met-F-AEA significantly inhibited, in tumors as well as transformed cells, the expression of the vascular endothelial growth factor, an angiogenetic factor known to be up-regulated by p21ras, as well as of one of its receptors, flt-1/VEGFR-1. The levels of the cyclin-dependent kinase inhibitor p27(kip1), which is down-regulated by p21ras, were instead increased by Met-F-AEA. All these effects were antagonized by the selective CB1 receptor antagonist SR141716A. Met-F-AEA inhibited in vitro the growth of a metastasis-derived thyroid cancer cell line more potently than a primary cancer cell line. Therefore, the hypothesis that CB1 receptor stimulation interferes not only with angiogenesis but also with metastatic processes was tested in a widely used model of metastatic infiltration in vivo, the Lewis lung carcinoma (3LL) in C57Bl/6 mice. Three weeks from the paw injection of 3LL cells, Met-F-AEA reduced significantly the number of metastatic nodes, in a way antagonized by SR141716A. Our findings indicate that CB1 receptor agonists might be used therapeutically to retard tumor growth in vivo by inhibiting at once tumor growth, angiogenesis, and metastasis. Key words: thyroid • cancer • endocannabinoid • p27 • lung • VEGF