青藤碱与顺铂协同诱导人纤维肉瘤HT-1080细胞凋亡

Synergistic induction of apoptosis by combination treatment of sinomenine with cisplatin in human fibrosarcoma HT-1080 cells

目的:探究青藤碱(sinomenine,SIN)对人纤维肉瘤HT-1080细胞顺铂敏感性的影响及其可能的分子机制。方法:采用CCK-8法检测青藤碱和顺铂对HT-1080细胞的毒性作用,采用Chou-Talalay中效分析法对两药的联合效应进行评价,采用流式细胞术检测细胞凋亡率,采用Western blot法检测铜离子转运蛋白1(copper transporter 1,CTR1)、谷胱甘肽S-转移酶π(glutathione S-transferase-π,GST-π)、Bcl-2和Bax的蛋白水平。结果:HT-1080细胞经顺铂或青藤碱处理48 h,细胞活力均受到显著抑制(P<0.05),IC50分别为6.50μmol/L和1.06 mmol/L,当抑制率超过25%时,二者联用呈协同效应;青藤碱与顺铂均可诱导HT-1080细胞凋亡(P<0.05),两药联合可使细胞凋亡率显著升高(P<0.05);青藤碱可上调CTR1和Bax的蛋白水平(P<0.05),下调GST-π和Bcl-2的蛋白水平(P<0.05)。结论:青藤碱可协同增强顺铂诱导的人纤维肉瘤HT-1080细胞凋亡,这可能与青藤碱上调CTR1和Bax蛋白水平以及下调GST-π和Bcl-2蛋白水平有关。

AIM:To investigate the synergistic antitumor effect of sinomenine(SIN)and cisplatin(DDP)against human fibrosarcoma HT-1080cells and its possible mechanism.METHODS:CCK-8assay was used to detect the cytotoxic effect of SIN and DDP in HT-1080cells,and the combination index(CI)of SIN and DDP was analyzed by Chou-Talalay method.The apoptosis was analyzed by flow cytometry.The protein levels of copper transporter1(CTR1),glutathione S-transferase-π(GST-π),Bcl-2and Bax were determined by Western blot.RESULTS:At48h after treatment with SIN or DDP at various concentrations,the cell viability was remarkably reduced(P<0.05),and the IC50values of DDP and SIN were6.50μmol/L and1.06mmol/L,respectively.When the inhibition rate of HT-1080cells exceeded25%,SIN in combination with DDP produced a synergistic effect(CI<1).Combination treatment of SIN with DDP signi-ficantly induced apoptosis of HT-1080cells.SIN up-regulated the protein levels of CTR1and Bax,and down-regulated the protein levels of GST-πand Bcl-2(P<0.05).CONCLUSION:SIN synergistic induces apoptosis with DDP in human fibrosarcoma HT-1080cells by up-regulating the protein levels of CTR1and Bax,and down-regulating the protein levels of GST-πand Bcl-2.