香豆素缩氨基酸型曼尼西碱香豆素类铜药物体内外抗肺癌增殖及其作用机制

Effect and their mechanism of action of copper （ n ） complexes with coumarin derivatives on lung cancer in vivo and vitro

目的研究香豆素缩氨基酸型曼尼西碱金属香豆素类铜药物（简称香豆素类铜药物）在体内外抗肺腺癌A549细胞的生物学活性。方法不同浓度香豆素类铜药物干预A549细胞，用MTT法、AnnexinV／PI及Westernblot进行检测体外抗肺癌活性。建立A549荷瘤裸鼠模型后，经过香豆素类铜药物的干预，绘制肿瘤生长曲线并计算肿瘤干预的抑瘤率，通过TUNEL法检测体内肿瘤细胞凋亡。结果香豆素类铜药物以浓度时间依赖性方式有效地抑制A549细胞增殖，抑制A549细胞活力的IC50为2．0μmol／L；香豆素类铜药物能以浓度依赖性方式诱导A549细胞凋亡（P〈0．05）；伴有caspase-3蛋白表达上调。香豆素类铜药物体内以浓度依赖性方式抑制肺癌A549移植瘤生长（P〈0．05），且凋亡明显增多（Pd0．05）。结论香豆素类铜药物对肺腺癌A549的体内外抑制生长作用均很明显，且可能通过caspase途径诱导细胞凋亡起作用。

Objective To investigate effect of the copper （ I ） complexes with coumarin derivatives （coumarin-copper drugs） on lung cancer cell （A549 cell line） in vivo and vitro and its mechanism of action. Methods A549 cells were treated with different concentrations of the coumarin-copper drugs, MTT assay was performed to determine cell ratio. Cell apoptosis was detected by Annexin V/PI flow cytometry. Western blot was performed to evaluate the protein expression of caspase-3. The subcutaneous transplantable tumor model of A549 cell in nude mice was established. The nude mice in each group received corresponding treated. The growth curves of the tumor were drawn and the tumor growth inhibition rates were calculated. The apoptosis index of subcutaneously transplanted tumor cells was served by TUNEL. Results Coumarin copper drugs effectively inhibited the proliferation of A549 in a dose-and-time dependent manner, and the IC50 was 2.0 μmol/L. Coumarin-copper drugs could induce the apoptosis of A549 cells in a dose dependent way （ P 〈0.05）, which was accompany with up- regulation of caspase-3. Coumarin-copper drugs effectively inhibited the growth of A549 tumor in a dose dependent manner, kept company with the apoptosis index of A549 tumor （ P 〈0.05）. Conclusions Coumarin-copper drugs may inhibit the proliferation of A549 cells through inducing cell apoptosis, which should be related to the up-regulation of easpase-3.