Japanese Journal of Aromatherapy, 16(1): (2015)

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1 Japan Journal of Aromatherapy Vol. 16, No. 1, T * 1, 2 Beneficial effects of essential oils on adult T-cell leukemia Abstract Adult T-cell leukemia ATL is a highly aggressive malignancy of CD4 T cells caused by human T cell leukemia virus type I HTLV-I. Transcription factor, NF-κB, is constitutively activated in primary ATL cells and plays a central role in leukemogenesis by contributing to the pathogenesis of ATL. Thus, specific NF-κB inhibitors block proliferation of ATL cells. Here we examined whether 30 essential oils exhibit NF-κB inhibitory effect as well as anti-atl effects using a human HTLV-I-infected T-cell line, MT-2. Among 30 oils, Geranium, Ylang-ylang, Juniper, and Jasmine oils significantly inhibited cellular growth. Geranium oil especially induced an inhibition of constitutive NF-κB activation as well as the caspase-dependent apoptotic cell death of MT-2. These results suggest that Geranium oil has strong NF-κB inhibitory and anti-atl effects. These findings demonstrate that Geranium oil and its components might be useful in molecular targeted therapies and palliative care for several cancer and inflammatory diseases, in which activation of NF-κB occurs. Key words: essential oil, cancer, adult T-cell leukemia, NF-κB, Geranium 1. T adult T-cell leukemia; ATL T I human T-lymphotropic virus type I; HTLV-I 1 HTLV-I 2 ATL ATL 3 HTLV-I T ATL NF-κB 4 NF-κB 1 NF-κB NF-κB NF-κB NF-κB ATL 5, 6 NF-κB NF-κB 7 NF-κB 8 13 NF-κB ATL Ryuichiro KIMURA* 1, 2 1. /Graduate School of Medicine, University of the Ryukyus 2. /Transdisciplinary Research Organization for Subtropics and Island Studies, University of the Ryukyus

2 16 Vol. 16, No. 1, 2015 NF-κB NF-κB IκB kinase IKK inhibitor of κb IκB -α IκB-α NF-κ B HTLV-I T Tax IKK NF-κB GAIA Excellent acute lymphoid leukemia; ALL Jurkat HTLV-I T MT Biological Industries RPMI 1640 Sigma-Aldrich 37, 5 CO 2 1. Cananga odorata Juniperus communis Pelargonium graveolens Melaleuca alternifolia Eucalyptus globulus Lavandula officinalis Citrus limon Rosmarinus officinalis Mentha piperita Citrus sinensis Salvia sclarea Citrus paradisi Santalum album Origanum majorana Citrus aurantium Boswellia carterii Citrus bergamia Rosa damascena 25 Styrax tonkinensis Piper nigrum Pogostemon cablin Cupressus sempervirens Vetiveria zizanioides Matricaria chamomilla Jasminum officinale Rosa damascena Commiphora myrrha Anthemis nobilis Cymbopogon flexuosus Melissa officinalis

3 Japan Journal of Aromatherapy Vol. 16, No. 1, MTT MTT Cell Counting Kit-8 DOJINDO 96 24, Cell Counting Kit-8 WST nm 680XR BIO-RAD 100 MTT 50 IC 50 v/v Microsoft Excel Z-VAD-FMK Promega triplicate cleaved caspase-8, phospho-iκb-α, IκB-α Cell Signaling Technology actin NeoMarkers Image J Actin 2.5 Apo2.7 Apo2.7 phycoerythrin PE Apo2.7 Beckman Coulter Epics XL Beckman Coulter Apo /7 8 9 Caspase-Glo 3/7 Assay, Caspase-Glo 8 Assay, Caspase-Glo 9 Assay Promega 1420 ARVO MX 2.7 Student t p ATL 30 HTLV-I T ALL NF-κB Jurkat HTLV-I T NF-κB MT-2 MTT Jurkat MT IC 50 v/v 2 Jurkat MT-2 2. Jurkat IC 50 v/v MT-2 IC 50 v/v

4 18 Vol. 16, No. 1, Jurkat MT-2 72 MTT 3 Jurkat MT-2 Student t *p 0.05, **p Jurkat MT-2 IC MT-2 Jurkat Jurkat 100 Jurkat 3 GAIA Excellent ATL MT-2 3 NF-κB 3.2 HTLV-I T 4 caspase MT caspase-8 6A 4

5 Japan Journal of Aromatherapy Vol. 16, No. 1, MTT 3 Jurkat MT-2 Student t *p 0.05, **p 0.01, ***p MTT 3 Jurkat MT-2 Student t *p 0.05, **p 0.01, ***p 0.001

6 20 Vol. 16, No. 1, GAIA Excellent MTT 3 Jurkat MT-2 Student t *p 0.05, **p 0.01, ***p NF-κB MT-2 24 Cleaved Caspase-8 Phospho-IκB-α Actin

7 Japan Journal of Aromatherapy Vol. 16, No. 1, A Z-VAD-FMK MT-2 24 MTT 3 Student t ***p B A Apo2.7 Apo2.7 C MT Student t **p 0.01 cleaved caspase-8 HTLV-I T 3.3 ATL NF- κ B 4 MT-2 NF-κB 1 NF-κB I κb-α IκB-α HTLV-I T MT-2 IκB-α MT-2 IκB-α NF-κB 6B MT-2 IκB-α IκB-α NF-κB NF-κB ATL 3.4 MT-2 24 MTT 7A 2 Z-VAD-FMK Apo2.7 7B Z-VAD-FMK

8 22 Vol. 16, No. 1, C 3/7 8 9 ATL 4. 4 ATL NF-κB NF-κB ATL NF-κB 7 ATL ATL NF-κB NF-κB 16 ATL NF-κB ATL NF-κB NFκB ATL NF-κB 16 NF-κB 17, 18 II 19 NF-κB 20 Geraniin NF-κB ATL NF-κB HTLV-I T ATL T Jurkat T Yoshida M., Miyoshi I., Hinuma Y.: Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease, Proc. Natl. Acad. Sci. USA, 79, Watanabe T.: HTLV-1-associated diseases, Int. J. He-

9 Japan Journal of Aromatherapy Vol. 16, No. 1, matol., 66, Matsuoka M., Jeang K. T.: Human T-cell leukaemia virus type 1 HTLV-1 infectivity and cellular transformation, Nat. Rev. Cancer, 7, Mori N., Fujii M., Ikeda S., Yamada Y., Tomonaga M., Ballard D. W., Yamamoto N.: Constitutive activation of NF-κB in primary adult T-cell leukemia cells, Blood, 93, Mori N., Yamada Y., Ikeda S., Yamasaki Y., Tsukasaki K., Tanaka Y., Tomonaga M., Yamamoto N., Fujii M.: Bay inhibits transcription factor NF-kappaB and induces apoptosis of HTLV-I-infected T-cell lines and primary adult T-cell leukemia cells, Blood, 100, Watanabe M., Ohsugi T., Shoda M., Ishida T., Aizawa S., Maruyama-Nagai M., Utsunomiya A., Koga S., Yamada Y., Kamihira S., Okayama A., Kikuchi H., Uozumi K., Yamaguchi K., Higashihara M., Umezawa K., Watanabe T., Horie R.: Dual targeting of transformed and untransformed HTLV-1-infected T cells by DHMEQ, a potent and selective inhibitor of NF-kappaB, as a strategy for chemoprevention and therapy of adult T-cell leukemia, Blood, 106, Olivier S., Robe P., Bours V.: Can NF-kappaB be a target for novel and efficient anti-cancer agents?, Biochem. Pharmacol., 72, Mori N., Ishikawa C., Uchihara J.-N., Yasumoto T.: Protein phosphatase 2A as a potential target for treatment of adult T cell leukemia, Curr. Cancer Drug Targets, 13, Tafuku S., Ishikawa C., Yasumoto T., Mori N.: Antineoplastic effects of fucoxanthin and its deacetylated product, fucoxanthinol, on Burkitt s and Hodgkin s lymphoma cells, Oncol. Rep., 28, Ishikawa C., Arbiser J. L., Mori N.: Honokiol induces cell cycle arrest and apoptosis via inhibition of survival signals in adult T-cell leukemia, Biochim. Biophys. Acta, 1820, Nakama S., Ishikawa C., Nakachi S., Mori N.: Antiadult T-cell leukemia effects of Bidens pilosa, Int. J. Oncol., 38, Tsumuraya T., Ishikawa C., Machijima Y., Nakachi S., Senba M., Tanaka J., Mori N.: Effects of hippuristanol, an inhibitor of eif4a, on adult T-cell leukemia, Biochem. Pharmacol., 81, Yamamoto K., Ishikawa C., Katano H., Yasumoto T., Mori N.: Fucoxanthin and its deacetylated product, fucoxanthinol, induce apoptosis of primary effusion lymphomas, Cancer Lett., 300, Miyoshi I., Kubonishi I., Yoshimoto S., Akagi T., Ohtsuki Y., Shiraishi Y., Nagata K., Hinuma Y.: Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells, Nature, 294, Kimura R., Ishikawa C., Rokkaku T., Janknecht R., Mori N.: Phosphorylated c-jun and Fra-1 induce matrix metalloproteinase-1 and thereby regulate invasion activity of 143B osteosarcoma cells, Biochim. Biophys. Acta, 1813, Shim J. U., Oh P. S., Lim K. T.: Anti-inflammatory activity of ethanol extract from Geranium sibiricum Linne, J. Ethnopharmacol., 126, Choi H. J., Choi H. J., Park M. J., Lee J. Y., Jeong S. I., Lee S., Kim K. H., Joo M., Jeong H. S., Kim J. E., Ha K. T.: The inhibitory effects of Geranium thunbergii on interferon-γ- and LPS-induced inflammatory responses are mediated by Nrf2 activation, Int. J. Mol. Med., 35, Maruyama N., Ishibashi H., Hu W., Morofuji S., Inouye S., Yamaguchi H., Abe S.: Suppression of carrageenan- and collagen II-induced inflammation in mice by geranium oil, Mediators Inflamm., 2006, Su Y. W., Chao S. H., Lee M. H., Ou T. Y., Tsai Y. C.: Inhibitory effects of citronellol and geraniol on nitric oxide and prostaglandin E2 production in macrophages, Planta Med., 76, Xiao F., Zhai Z., Jiang C., Liu X., Li H., Qu X., Ouyang Z., Fan Q., Tang T., Qin A., Gu D.: Geraniin suppresses RANKL-induced osteoclastogenesis in vitro and ameliorates wear particle-induced osteolysis in mouse model, Exp. Cell. Res., 330,

10 24 Vol. 16, No. 1, 2015 T ATL T I HTLV-I ATL NF-κB NF-κB ATL ATL NF-κB HTLV-I T 30 4 HTLV-I T NF-κB NF-κB ATL NF-κB T NF-κB TEL: FAX: kimuraryuichiro@yahoo.co.jp

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text-H1_ol Ylang Ylang Cananga odorata Orange Citrus sinensis Citrus aurantium Chamomile.G Matricaria recutita Chamomile.R Chamaemelum nobile Cardamon Elettaria cardamomum Clary Sage Salvia sclarea Grapefruit Citrus