欧前胡素增强多柔比星对 HeLa 细胞的抗肿瘤效应

摘要

目的：研究欧前胡素是否能提高宫颈癌HeLa细胞株对多柔比星的敏感性。方法： MTT法检测HeLa细胞用欧前胡素和多柔比星处理后的活力。 Western blot检测HeLa细胞用欧前胡素和多柔比星处理后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HeLa细胞用欧前胡素和多柔比星处理后的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多柔比星治疗宫颈癌效果的影响。结果：欧前胡素在体外可显著提高多柔比星对宫颈癌细胞系HeLa的杀伤活性。欧前胡素可显著降低HeLa细胞Mcl－1的表达，而多柔比星对Mcl－1的表达水平无影响。相比于欧前胡素或多柔比星单治疗组，两者联合可显著诱导HeLa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多柔比星联合欧前胡素对HeLa细胞的杀伤活性。结论：欧前胡素通过靶向于Mcl－1增强多柔比星对宫颈癌细胞的杀伤活性。%AIM:To determine whether imperatorin would enhance the effect of doxorubicin therapy on cervi -cal cancer in vitro.METHODS:The viability of HeLa cells treated with imperatorin and doxorubicin was determined by MTT assay in vitro.The expression of Bcl-2 protein family ( Mcl-1, Bcl-2, Bcl-xL, Bad and Bax ) in HeLa cells treated with imperatorin and doxorubicin was evaluated by Western blot analysis .The apoptosis and mitochondrial membrane poten-tial (ΔΨm ) in the HeLa cells treated with imperatorin and doxorubicin were analyzed by flow cytometry .A Mcl-1 expres-sion vector was constructed , and its role in the cytotoxicity of imperatorin plus doxorubicin to HeLa cells was detected by MTT assay.RESULTS: Addition of imperatorin significantly enhanced the cytotoxicity of doxorubicin to HeLa cells in vitro.Mcl-1 expression was down-regulated by imperatorin but was not influenced by doxorubicin in the HeLa cells .A com-bination of imperatorin and doxorubicin induced apoptosis and ΔΨm collapse more significantly compared with the treatment with imperatorin or doxorubicin alone .Furthermore, the imperatorin-induced sensitization for doxorubicin cytotoxicity to HeLa cells was abolished by the transfection with Mcl-1 expression plasmid .CONCLUSION:The combination of doxoru-bicin with imperatorin enhances the antitumor effect of doxorubicin on cervical cancer cells via targeting Mcl -1.