欧前胡素对顺铂的骨肉瘤细胞抗肿瘤作用的增效作用

Anti-tumor Effect and Mechanism of Imperatorin Enhances the Cytotoxicity of Cisplatin Osteosarcoma Cells

关键词:  欧前胡素  顺铂  Mcl-1  骨肉瘤

Key words:  imperatorin  cisplatin  Mcl-1  osteosarcoma

摘要:

目的 研究欧前胡素是否能提高人骨肉瘤细胞(human osteosarcoma cells，HOS细胞)对顺铂的敏感性。方法 HOS细胞用欧前胡素和顺铂处理，MTT法检测体外细胞活力，Western blot检测Bcl-2 蛋白家族成员(Mcl-1、Bcl-2、Bcl-xl、Bad和Bax)的表达水平，流式细胞术检测细胞凋亡水平和线粒体膜电位的变化情况。构建Mcl-1真核表达载体，MTT法检测Mcl-1表达载体转染对欧前胡素联合顺铂治疗骨肉瘤疗效的影响。结果 欧前胡素在体外可显著提高顺铂对骨肉瘤细胞系HOS的杀伤活性。欧前胡素可显著降低HOS细胞Mcl-1的表达，而顺铂对Mcl-1的表达水平无影响。相比于欧前胡素或顺铂单治疗组，两者联合可显著诱导HOS细胞发生凋亡并降低其线粒体膜电位。体外转染Mcl-1真核表达载体显著降低顺铂联合欧前胡素对HOS细胞的杀伤活性。结论 欧前胡素通过靶向于Mcl-1增强顺铂对骨肉瘤细胞的杀伤活性。

Abstract:

OBJECTIVE To study the effect of the addition of imperatorin in cisplatin therapy in osteosarcoma(HOS cells) in vitro. METHODS HOS cells were treated with imperatorin and cisplatin, and then their viability was determined by MTT assay in vitro, the expression of Bcl-2 protein family (Mcl-1, Bcl-2, Bcl-xl, Bad and Bax) was evaluating by western blot, and flow cytometry was used for apoptosis detection and measurement of mitochondrial membrane potential. Construct Mcl-1 expression vector, and then detected the role of Mcl-1 vector toward imperatorin plus cisplatin–inducing cytotoxicity in HOS cells by MTT assay. RESULTS Addition of imperatorin significantly enhanced the cytotoxicity of cisplatin to HOS cells in vitro. Mcl-1 expression was down-regulated by imperatorin but not influenced by cisplatin in HOS cells. A combination of imperatorin and cisplatin induced apoptosis and mitochondrial membrane potential collapse more significantly compared with treatment of imperatorin or cisplatin alone. Furthermore, the imperatorin-induced sensitization for cisplatin-cytotoxicity to HOS cells was abolished by the transfection of Mcl-1 expression plasmid. CONCLUSION Combination with imperatorin can enhance the anti-tumor activity of cisplatin to osteosarcoma cells via targeting Mcl-1.