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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.

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