Tumor metastasis is a major cause of death in cancer patients, and its blockade is believed to enable cancer patients to survive. 1) Thus, it is important to find promising agents with anti-metastatic activity.One key event in the formation of tumor metastasis is cell motility. A number of host-derived factors have been shown to modulate the motile activity of tumor cells. Hepatocyte growth factor (HGF) is one such factor and has been shown to stimulate the invasion and metastasis as well as migration in a variety of tumor cells.2-4) Clinical investigations also indicate the importance of HGF in tumor metastasis by the findings that a significantly elevated serum level of HGF is observed in the patients with lung, 5) prostate, 6) gastric, 7) and breast cancer. 8) From these observations, regulation of HGF activity is recognized as a possible step in tumor metastasis therapy, however, little has been reported about compounds having anti-HGF activity.We recently reported that evodiamine has remarkable inhibitory activity on in vitro invasion and migration of tumor cells.9,10) Evodiamine is one of the main constituents of Evodiae Fructus 11) and has been shown to possess anti-tumor growth, 12,13) properties. With regard to the growth inhibition by evodiamine, Fei et al. have clearly demonstrated the molecular mechanisms by which evodiamine increases the expression of the apoptosis inducer Bax and decreases the apoptosis suppressor Bcl-2 in mitochondria, triggering the activation of caspase-3 and DNA fragmentation. 13) Moreover, the existence of unknown apoptotic pathways is speculated in the evodiamine-induced apoptosis. In contrast, no reports has been seen on the effects of evodiamine on HGF activity.In the present study, we investigated the effects of evodiamine on HGF-induced invasion and migration of tumor cell lines, colon 26-L5 carcinoma, B16-F10 melanoma, and Lewis lung carcinoma.
MATERIALS AND METHODS
MaterialsEvodiamine and hepatocyte growth factor (HGF) were purchased from Matsuura Yakugyo Co., Ltd. (Aichi, Japan) and Funakoshi Co., Ltd. (Tokyo, Japan), respectively. Evodiamine and HGF were dissolved in dimethyl sulfoxide (DMSO) and distilled water, respectively. In each in vitro treatment, evodiamine was diluted with DMSO, and then the solution was added into cell-suspended medium to make a final concentration of 1% DMSO.Cells and Cell Culture Murine colon 26-L5 adenocarcinoma was kindly provided by Prof. I. Saiki (Toyama Med. Pharm. Univ., Inst. of Natural Medicine, Toyama, Japan). Murine B16-F10 melanoma was kindly provided by Dr. S. Wakuzawa (Hokuriku Univ., Kanazawa, Japan). Lewis lung carcinoma (LLC) was purchased from the RIKEN Cell Bank. Colon 26-L5 cells and B16-F10 cells were maintained in RPMI-1640 supplemented with 10% fetal calf serum (FCS), 2-mercaptoethanol, 100 U/ml penicillin, and 0.1 mg/ml streptomycin. LLC was maintained in DMEM with 10% FCS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin.Cell Invasion and Migration Assay Tumor cell invasion through a reconstituted basement membrane ...