antitumor activity and macrophage nitric oxide producing action of medicinal herb, crassocephalum crepidioides抗肿瘤活性和巨噬细胞一氧化氮产生行动的草药,crassocephalum crepidioides.pdfVIP

Tomimori et al. BMC Complementary and Alternative Medicine 2012, 12:78 /1472-6882/12/78 RESEARCH ARTICLE Open Access Antitumor activity and macrophage nitric oxide producing action of medicinal herb, Crassocephalum crepidioides 1,2† 1,3† 1,4 1 1,4 1* Koh Tomimori , Shinji Nakama , Ryuichiro Kimura , Kazumi Tamaki , Chie Ishikawa and Naoki Mori 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