【摘要】 目的:以人乳腺癌细胞MCF-7和耐多柔比星(doxorubicin,DOX)的人乳腺癌细胞MCF-7/DOX为研究对象,观察新型多芳烯姜黄素衍生物T63对这两株细胞系的影响,探讨T63能否逆转MCF-7/DOX细胞对多柔比星(DOX)的耐药性。方法:用MTT法检测T63的细胞毒性,同时检测T63对细胞株MCF-7/DOX药物敏感性的影响。用流式细胞术PI单染法分析T63或T63联合DOX对细胞周期的影响,观察细胞凋亡情况。用Western blot检测细胞凋亡相关蛋白表达。用Real-time PCR法检测相关基因的转录情况。结果:T63能够有效抑制MCF-7和MCF-7/DOX的增殖,且药效强于姜黄素,而T63对MCF-7和MCF-7/DOX的抑制增殖的效应差异不大。使用非细胞毒性剂量(0.75μmol/L)的T63能有效提高MCF-7/DOX对多柔比星的敏感性。PI单染法显示单独使用T63或者非细胞毒性剂量T63与多柔比星联合使用均能促进细胞凋亡。Western blot结果显示T63能够上调p53、p21、p27、Bim、Bax的表达,下调Bcl-2的表达。Real-time PCR结果表明T63对p53无显著影响。结论:T63可通过促进细胞凋亡的方式逆转MCF-7/DOX对多柔比星的耐药性,从而提示其可能具有临床应用价值,其具体分子机制有待进一步研究。更多还原

【Abstract】 Objective:To investigate the reversion of MCF-7/DOX with T63 in vitro.Methods:Cytotoxicity of doxorubicin(DOX),DOX plus T63 were evaluated by measuring the inhibition of cell growth by MTT assay.Flow cytometry(FCM) was performed to determine cell cytolysis and apoptosis.The expression of apoptosis related proteins were determined by Western blot,and the transcriptional levels of related genes were determined by real-time PCR.Results:Cytotoxicity assays showed that T63 significantly inhibited cell growth.The inhibition effects of T63 were greater than that of curcumin.Apoptosis assays showed T63 potentiate DOX cytotoxicity and restore chemotherapy sensitivity in the MCF-7/DOX.Western blot showed that T63 up-regulated the expression of p53,p21,p27,Bim,Bax and down-regulated the expression of Bcl-2.The results of real-time PCR showed that T63 had limited influence on mRNA levels of p53.Conclusion:These results provided a biochemical basis for possible clinical application of T63 in combination with doxorubicin in the treatment of chemo-resistance tumors,but the detailed mechanisms are needed further investigation.更多还原