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Open Access Highly Accessed Research article

Antitumor activity and macrophage nitric oxide producing action of medicinal herb, Crassocephalum crepidioides

Koh Tomimori12, Shinji Nakama13, Ryuichiro Kimura14, Kazumi Tamaki1, Chie Ishikawa14 and Naoki Mori1*

Author Affiliations

1 Department of Microbiology and Oncology, Graduate School of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa, 903-0215, Japan

2 Department of Psychiatry, Naha City Hospital, 2-31-1 Furujima Naha, Okinawa, 902-8511, Japan

3 Musashino Research Institute for Immunity, 790 Nishizatozoe, Gusukube, Miyako Island, Okinawa, 906-0106, Japan

4 Transdisciplinary Research Organization for Subtropics and Island Studies, 1 Senbaru Nishihara, Okinawa, 903-0213, Japan

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BMC Complementary and Alternative Medicine 2012, 12:78  doi:10.1186/1472-6882-12-78

Published: 21 June 2012

Abstract

Background

Crassocephalum crepidioides, a plant distributed in Okinawa Islands, is known in folk medicine; however, its anticancer activity has not been investigated. The aim of this study was to determine the in vitro and in vivo antitumor activities of C. crepidioides on murine Sarcoma 180 (S-180) and related molecular mechanisms.

Methods

The antitumor effect of C. crepidioides was evaluated in S-180-cell-bearing mice. Cell growth was assessed using a colorimetric assay. Nitrite and nitrate levels were measured by colorimetry. The expression levels of inducible NO synthase (iNOS) in murine RAW264.7 macrophages was assessed by reverse transcriptase-polymerase chain reaction. Activation of iNOS promoter was detected by reporter gene. Activation of nuclear factor-κB (NF-κB) was evaluated by electrophoretic mobility shift assay. The role of NF-κB signaling was analyzed using inhibitors of NF-κB and dominant-negative mutants, and Western blot analysis.

Results

C. crepidioides extract delayed tumor growth in S-180-bearing mice. However, it did not inhibit S-180 cell growth in vitro. Supernatant of cultured C. crepidioides-stimulated RAW264.7 macrophages was cytotoxic to S-180 cells. This cytotoxicity was associated with nitric oxide (NO) production. NF-κB signaling pathway was crucial for the transcriptional activation of iNOS gene. Isochlorogenic acid, a component of C. crepidioides, induced NF-κB activation and iNOS expression.

Conclusions

The results highlight the oncolytic and immunopotentiation properties of C. crepidioides mediated through NF-κB-induced release of NO from macrophages.