摘要Objective:To evaluate anti-melanoma effect of ethanol extract of Ilex hainanensis Merr.(IME)and elucidate its underlying mechanism.Methods:Thirty-six tumor-bearing mice were randomized into 6 groups (n=6) as follows:model group,IME 25,50,100,and 200 mg/kg groups and dacarbazine (DTIC) 70 mg/kg group.The mice in the IME treatment groups were intragastdcally administered with IME 25,50,100 or 200 mg/kg per day,respectively.The mice in the DTIC group were intrapedtoneally injected with DTIC 70 mg/kg every 2 days.The drug administration was lasting for 14 days.The cell viability was evaluated by 3-(4,5-dime-thylthylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay.Flow cytometry was employed to detect cell cycle and apoptosis.The gene and protein expressions of nuclear factor κ B-p65 (NF-κ B-p65),Bcl-2,B-cell lymphomaextra large (Bcl-xL) and Bax were detected by quantitative real-time polymerase chain reaction and Westem blot analyses.Caspases-3,-8,and-9 activities were detected using the colodmetdc method.In addition,a B16-F10 melanoma xenograft mouse model was used to evaluate the anti-cancer activity of IME in vivo.Furthermore,a survival experiment of tumor-bearing mice was also performed to evaluate the possible toxicity of IME.Resalto:IME significantly inhibited the proliferation of B16-F10 cells (P＜0.01).Flow cytometric analysis showed that IME induced G1/S cell cycle arrest and apoptosis (both P＜0.01).IME inhibited activation of NF-κ B,decreased the gene and protein expressions of Bcl-2,Bcl-xL,and increased the gene and protein expressions of Bax (all P＜0.01).In addition,IME induced the activation of Caspases-3,-8,and-9 in B16-F10 cells.The study in vivo showed that IME significantly reduced tumor volume (P＜0.01),and the inhibitory rate came up to 68.62％.IME also induced large areas of necrosis and intra-tumoral apoptosis that correlated with a reduction in tumor volume.Survival experiment showed that treatment with IME for 14 days significantly prolonged survival time and 20％ of mice in the IME 200 mg/kg group were still alive until the 50th day.Notably,IME showed no apparent side-effects dudng the treatment pedod.Coaclusion:IME exhibited significant anti-melanoma activity in vftro and in vivo,suggesting that IME might be a promising effective candidate with lower toxic for malignant melanoma therapy.