目的：观察新型香豆素类酰腙—铜配合物（以下缩写为CCCD）在体内外对肺癌A549细胞的抑制作用，并探讨其机制。方法培养肺癌A549细胞，分别加入5、10、20、30、50、80、120、160μmol／L的CCCD，干预72 h后，采用MTT法，计算细胞生长抑制率（ IR ）。将A549细胞分为干预组、对照组，干预组分别加入10、20、40μmol／L CCCD，对照组加入PBS，采用流式细胞术检测各组细胞凋亡情况并计算细胞凋亡率，采用Western blot法检测各组细胞Caspase-3蛋白表达。取18只裸鼠建立肺癌荷瘤鼠模型，分为观察1组、观察2组、对照组，每组各6只，分别予尾静脉注射4、8 mg／kg CCCD及PBS，1次／周，共干预3周，干预结束后测算各组肿瘤体积并计算抑瘤率。随后处死各组裸鼠，取瘤体组织，应用TUNEL法检测各组肿瘤细胞凋亡情况并计算凋亡指数（ AD）。结果加入5、10、20、30、50、80、120、160μmol／L CCCD 后， A549细胞 IR 分别为8．80％、16．52％、37．24％、55．75％、77．22％、87．16％、95．25％、98．70％，随着药物浓度增高，IR呈增高趋势。干预组加入10、20、40μmol／L CCCD后，细胞凋亡率均高于对照组（P均＜0．05）。干预组Caspase-3蛋白表达高于对照组（P＜0．05）。观察1组、观察2组、对照组AD分别为16．83％±8．44％、24．65％±11．24％、3．30％±2．12％，各组间比较P均＜0．05。观察1组、观察2组抑瘤率分别为51．08％、56．78％。结论 CCCD在体内外均可抑制肺癌A549细胞的生长，促进细胞凋亡，其作用机制可能与经Caspase-3途径诱导细胞凋亡有关。%Objective To observe the inhibitory effect of a new copper (Ⅱ) complex with coumarin derivatives ( CCCD) on lung cancer cell line A549 in vivo and in vitro and to investigate the mechanism .Methods The lung cancer A549 cells were cultured and were treated with 5, 10, 20, 30, 50, 80, 120, 160μmol/L CCCD for 72 h, then the inhibi-tion rate ( IR) of tumor cells was calculated by using MTT assay .A549 cells were divided into the intervention group which was treated with 10 , 20 and 40μmol/L CCCD and the control group which was treated with PBS .The apoptosis and the ap-optosis rate were analyzed by flow cytometry , and the expression of caspase-3 was detected by Western blotting .Further-more, 18 tumor-bearing nude mice were divided into the observation group 1, observation group 2 and the control group , which were treated with 4, 8 mg/kg CCCD and PBS once a week for three weeks , and then they were executed to detect the tumor volume and tumor inhibition rate .After that, the tumor tissues were obtained to detect the apoptosis and apoptosis in-dex (AD) by TUNEL method.Results The inhibition rates of A549 cells were 8.80%, 16.52%, 37.24%, 55.75%, 77.22%, 87.16%, 95.25%and 98.70%after being treated with 5, 10, 20, 30, 50, 80, 120 and 160μmol/L CCCD. The IR showed an increased tendency with the increased concentration of the drugs .And being after treated with 10, 20 and 40 μmol/L CCCD, the apoptosis rate and expression level of caspase-3 in the intervention group were significantly higher than those of the control group (all P<0.05).The AD in the observation groups 1, 2 and the control group were 16.83%±8.44%, 24.65%±11.24%and 3.30%±2.12%(all P<0.05).And the tumor inhibition rates in the observation groups 1 and 2 were 51.80%and 56.78%, respectively.Conclusions CCCD may inhibit the growth of A549 cells in vi-vo and in vitro and enhance the apoptosis , and its mechanism might be related to inducing tumor cell apoptosis through acti-vating caspase-3.
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