Cannabidiol Induces Cytotoxicity and Cell Death via Apoptotic Pathway in Cancer Cell Lines

  • ChoiPark, Won-HyungHyun-Do (Department of Medical Zoology, College of Medicine, KyungHee UniversityDivision of Anatomy & Developmental Biology, Department of Oral Biology, BK21 Project, College of Dentistry, Yonsei University) ;
  • Baek, Seung-Hwa (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University) ;
  • Chu, Jong-Phil (Department of Medical Zoology, College of Medicine, KyungHee University) ;
  • Kang, Mae-Hwa (Department of Medical Zoology, College of Medicine, KyungHee University) ;
  • Mi, Yu-Jing (Department of Medical Zoology, College of Medicine, KyungHee University) ;
  • Published : 2008.06.30


In view of obtaining potential anticancer compounds, we studied the inhibitory activity and the cytotoxic effects of a candidate compound in cancer cells. The cytotoxic effects of cannabidiol (CBD) in vitro were evaluated in NIH3T3 fibroblasts, B16 melanoma cells, A549 lung cancer cells, MDA-MB-231 breast cancer cells, Lenca kidney cells and SNU-C4 colon cancer cells. The cells were cultured in various concentrations of CBD for 48 h and 25 ${\mu}$M of CBD for 6-36 h. The cells were observed to exhibit inhibitory effects of the cell viability in their growth, and then cytotoxicity was estimated. The inhibitory activity of CBD was increased in all cancer cells and showed especially strong increment in breast cancer cells. The cytotoxicity of CBD increased in a dose- and time-dependent manner with growth inhibition in all cancer cell lines. Also, to assess the membrane toxicity induced by CBD, we investigated lactate dehydrogenase (LDH) release. After treatment with various concentrations of CBD, LDH release rate of cancer cells was accelerated. On the other hand, in the induction of cell death, caspase-3, -8 and -9 activations were detected in cancer cells after treatment with various concentrations of CBD, and CBD effectively induced activity of caspase-3, -8 and -9 in A549 lung cancer cells, MDAMB-231 breast cancer cells and Renca kidney cells. Therefore these results suggest that CBD has a possibility of anticancer agents and anticancer effects against cancer cells by modulation of apoptotic pathway in the range of 5-80 ${\mu}$M concentration.


  1. Andersson, D., Liu, J. J., Nilsson, A. and Duan, R. D. (2003). Ursolic acid inhibits proliferation and stimulates apoptosis in HT29 cells following activation of alkaline sphingomyelinase. Anticancer Res. 23, 3317-3322
  2. Asakawa, Y. (1995). 'In Progress in the Chemistry of Organic Natural Products'; Herz, W., Kirby, G. W., Moore, R. E., Steglich, W. and Tamm, C. eds. Springer: Vienna, Vol : 65
  3. Bold, R. J., Termuhlen, P. M. and McConkey, D. J. (1997). Apoptosis, cancer and cancer therapy. Surg Oncol. 6, 133-142
  4. Buja, L. M., Eigenbrodt, M. L. and Eigenbrodt, E. H. (1993). Apoptosis and necrosis. Basic types and mechanisms of cell death. Arch Pathol Lab Med. 117, 1208-1214
  5. Buttke, T. M. and Sandstrom, P. A. (1994). Oxidative stress as a mediator of apoptosis. Immunol Today. 15, 7-10
  6. Carrier, E. J., Auchampach, J. A., and Hillard, C. J. (2006). Inhibition of an equilibrative nucleoside transporter by cannabidiol: a mechanism of cannabinoid immuno-suppression. Proc Natl Acad Sci U S A. 103, 7895-7900
  7. Casanova, M. L., Blazquez, C., Martinez-Palacio, J., Villanueva, C., Fernandez-Acenero, M. J., Huffman, J. W., Jorcano, J. L. and Guzman, M. (2003). Inhibition of skin tumor growth and angiogenesis in vivo by activation of cannabinoid receptors. J Clin Invest. 111, 43-50
  8. Chiang, L. C., Ng, L. T., Chiang, W., Chang, M. Y. and Lin, C. C. (2003). Immunomodulatory activities of flavonoids, monoterpenoids, triterpenoids, iridoid glycosides and phenolic compounds of Plantago species. Planta Medica. 69, 600-604
  9. Consroe, P., Martin, A. and Singh, V. (1981). Antiepileptic potential of cannabidiol analogs. J. Clin. Pharmacol. 21, 428-436
  10. Consroe, P., Benedito, M. A., Leite, J. R. and Carlini, E. A., Mechoulam R. (1982). Eur J Pharmacol. 83, 293-298
  11. Esposito, G., De Filippis, D., Maiuri, M. C., De Stefano, D., Carnuccio, R. and Iuvone, T. (2006). Cannabidiol inhibits inducible nitric oxide synthase protein expression and nitric oxide production in beta-amyoid stimulated PC12 neurons through p38 MAP kinase and NF-kappaB involvement. Neurosci Lett. 399, 91-95
  12. Guimaraes, F. S., Chiaretti, T. M., Graeff, F. G. and Zuardi, A. W. (1990). Anti-anxiety effect of cannabidiol in the elevated plusmaze. Psychopharmacol (Berl). 100, 558-559
  13. Garcia-Arencibia, M., Gonzalez, S., De Lago, E., Ramos, J. A., Mechoulam, R. and Fernandez-Ruiz, J. (2007). Evaluation of the neuroprotective effect of cannabinoids in a rat model of Parkinson's disease: importance of antioxidant and cannabinoid receptor-independent properties. Brain Res. 1134, 162- 170
  14. Giudice, E. D., Rinaldi, L., Passarotto, M., Facchinetti, F., D'Arrigo, A., Guiotto, A., Carbonare, M. D., Battistin, L. and Leon, A. (2007). Cannabidiol, unlike synthetic cannabinoids, triggers activation of RBL-2H3 mast cells. J Leukoc Biol. 81, 1512-1522
  15. Grub, S., Persohn, E., Trommer, W. E. and Wolf, A. (2000). Mechanisms of cyclosporine A-induced apoptosis in rat hepatocyte primary cultures. Toxicol Appl Pharmacol. 163, 209- 220
  16. Hah, J. C., Rhew, T. H., Choi, E. S., Chung, H. Y. and Park, K. Y. (1992). Anti-tumor effect of ursolic acid against inbred hepatoma in CBA/J Mouse. Journal of Korean Cancer Association. 24, 790-794
  17. Harmand, P. O., Duval, R., Liagre, B., Jayat-Vignoles, C., Beneytout, J. L., Delage, C. and Simon, A. (2003). Ursolic acid induces apoptosis through caspase-3 activation and cell cycle arrest in HaCat cells. Int J Oncol. 23, 105-112
  18. Kerr, J. F. R., Wyllie, A. H. and Currie, A. R. (1972). Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics. British Journal of Cancer. 26, 239-257
  19. Lee, K. H., Lim, Y. M., Wu, T. S., Zhang, D. C., Yamagishi T., Hayashi T., Hall I. H., Chang J. J. and Wu R. Y. (1988). The cytotoxic principles of Prunella vulgaris, Psychotria serpens, and Hyptis capitata: ursolic acid and related derivatives. Planta Medica. 54, 308-311
  20. Ligresti, A., Moriello, A. S., Starowicz, K., Matias, I., Pisanti, S., De petrocellis, L., Laezza, C., Portella, G., Bifulco, M. and Di Marzo, V. (2006). Antitumor activity of plant cannabinoid with emphasis on the effect of cannabidiol on human breast carcinoma. J Pharmacol Exp Ther. 318, 1375-1387
  21. Martin, B. R. (1986). Cellular effects of cannabinoids. Pharmacol Rev. 38, 45-74
  22. Martin, A. R., Consroe, P., Kane, V. V., Shah, V., Singh, V., Lander, N., Mechoulam, R. and Srebnik, M. (1987). Structureanticonvulsant activity relationships of cannabidiol analogs. NIDA Res Monogr. 79, 48-58
  23. Mckallip, R. J., Jia, W., Schlomer, J., Warren, J. W., Nagarkatti, P. S. and Nagarkatti, M. (2006). Cannabidiol-induced apoptosis in human leukemia cells: A novel role of cannabidiol in the regulation of p22phox and Nox4 expression. Mol Pharmacol. 70, 897-908
  24. Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 65, 55-63
  25. Nicoletti, I., Migliorati, G., Pagliacci, M. C., Grignani, F. and Riccardi C. (1991). A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 139, 271-279
  26. Nicholson, D. W., Ali, A., Thomberry, N. A., Vaillancourt, J. P., Ding, C. D., Gallant, M., Gareau, Y., Griffin, P. R., Labelle, M. and Lazebnik, Y. A. (1995). Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature. 376, 37-43
  27. Perry, N. B., Forster, L. M., Lorimer, S. D., May, B. C. and Weavers, R. T. (1996). Isoprenyl phenyl ethers from liverworts of the genus Trichocolea: cytotoxic activity, structural corrections, and synthesis. J. Nat. Prod. 59, 729-733
  28. Pertwee, R. G. (1997). Pharmacology of cannabinoid CB1 and CB2 receptors. Pharmacol Ther. 74, 129-180
  29. Shishodia, S., Majumdar, S., Banerjee, S. and Aggarwal, B. B. (2003). Ursolic acid inhibits nuclear factor-kappaB activation induced by carcinogenic agents through suppression of IkappaBalpha kinase and p65 phosphorylation: correlation with down-regulation of cyclooxygenase 2, matrix metalloproteinase 9, and cyclin D1. Cancer Res. 63, 4375-4383
  30. Soo, L. Y., Jin, D. Q., Beak, S. M., Lee, E. S. and Kim, J. A. (2003). Inhibition of ultraviolet-A-modulated signaling pathways by asiatic acid and ursolic acid in HaCaT human keratinocytes. Eur J Pharmacol. 476, 173-178
  31. Vaccani, A., Massi, P., Colombo, A., Rubino, T. and Parolaro, D. (2005). Cannabidiol inhibits human glioma cell migration through a cannabinoid receptor-independent mechanism. Br J Pharmacol. 144, 1032-1036
  32. Weiss, L., Zeira, M., Reich, S., Har-Noy, M., Mechoulam, R., Slavin, S. and Gallily, R. (2006). Cannabidiol lowers incidence of diabetes in non-obese diabetic mice. Autoimmunity. 39, 143-151
  33. Wyllie, A. H. (1992). Apoptosis and the regulation of cell numbers in normal and neoplastic tissues: an overview. Cancer Metastasis Rev. 11, 95-103
  34. Wyllie, A. H. (1985). The biology of cell death in tumours. Anticancer Res. 5, 131-136
  35. Wyllie, A. H., Kerr, J. F. and Currie, A. R. (1980). Cell death: the significance of apoptosis. Int Rev Cytol. 68, 251-306

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