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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。
Objective To investigate whether imperatorin can increase the sensitivity of human osteosarcoma cells to cisplatin. Methods HOS cells were treated with imperatorin and cisplatin. The viability of cells in vitro was detected by MTT assay. The expression of Bcl-2, Bcl-2, Bcl-2, Bad and Bax was detected by Western blot. Flow cytometry was used to detect the changes of apoptosis and mitochondrial membrane potential. Construction of Mcl-1 eukaryotic expression vector, MTT assay Mcl-1 expression vector transfection of imperatorin and cisplatin treatment of osteosarcoma effect. Results Imperatorin can significantly increase the killing activity of cisplatin on human osteosarcoma cell line HOS in vitro. Imperatorin significantly reduced Mcl-1 expression in HOS cells, whereas cisplatin had no effect on Mcl-1 expression. Compared with imperatorin or cisplatin monotherapy group, the combination of the two can significantly induce apoptosis of HOS cells and reduce their mitochondrial membrane potential. In vitro transfection of Mcl-1 eukaryotic expression vector significantly reduced the killing activity of cisplatin combined with imperatorin on HOS cells. Conclusion Imperatorin enhances the killing activity of cisplatin on osteosarcoma cells by targeting Mcl-1.