钙调素拮抗剂O-(4-乙氧基)-丁基-小檗胺增强阿霉素杀伤MCF-7/ADR细胞的机制研究  

Mechanism of synergistic antitumor effects of EBB and doxorubicin on multidrug resistant MCF-7/ADR

作　　者：赵英新[1] 刘荣[1] 范冬梅[1] 任思楣[1] 李崴[1] 师锐赞[1] 张砚君[1] 杨铭[1] 

机构地区：[1]中国医学科学院北京协和医学院血液病医院血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中国药理学通报》2010年第2期231-235,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30400558);津市应用基础研究重点资助项目(No07JCZDJC04900)

摘　　要：目的研究钙调素拮抗剂0-4-乙氧基-丁基-小檗胺(EBB)增强阿霉素诱导乳腺癌多药耐药细胞系MCF-7/ADR细胞的杀伤作用及其相关机制。方法用MTT法测定阿霉素、EBB单独及联合用药对阿霉素杀伤乳腺癌多药耐药细胞系(MCF-7/ADR)及其亲代细胞系(MCF-7)的作用的IC50值,用不同浓度EBB处理MCF-7/ADR细胞后用FACS法分析EBB对阿霉素诱导细胞凋亡及对mdr1mRNA和P-gp蛋白水平表达的影响,通过激光共聚焦显微镜观察EBB处理前后及用EBB预处理24和48h后MCF-7/ADR和MCF-7细胞内阿霉素浓度的改变。结果MTT结果显示EBB对MCF-7和MCF-7/ADR都具有抗肿瘤活性;EBB还能协同提高阿霉素的细胞毒作用,MCF-7组两药相互作用指数(CDI)值为0.73,MCF-7/ADR组CDI值为0.49,其对耐药细胞的协同作用更为明显。随EBB剂量增加,低剂量阿霉素诱导MCF-7/ADR细胞凋亡增加而且P-gp蛋白表达水平逐渐下降,细胞内阿霉素浓度逐渐提高,而且用EBB预处理MCF-7/ADR细胞24和48h后细胞内阿霉素和罗丹明浓度也逐渐提高。结论EBB是有效的肿瘤细胞化疗药物,它不但能直接抑制P-gp功能还具有下调P-gp蛋白表达的作用,从而有效逆转MCF-7/ADR细胞的耐药现象,协同增强化疗药物对耐药细胞的杀伤作用。Aim To study the mechanism of synergistic antitumor of EBB and doxorubicin in doxoruhicin-resistant MCF-7/ADR breast carcinoma cells. Methods The antitumor activity of doxorubiein alone and its combination with EBB were measured by MTT assay in MCF-7/ADR and MCF-7cells. The rate of doxorubicin-induced apoptosis and the protein and mRNA levels of P-glycoprotein （P-gp） were determined in MCF-7/ ADR treated with EBB by flow cytometry （FACS）, respectively. Laser scanning confocal microscopy was used to detect the intracellular accumulation of drug in EBB-treated MCF-7 and MCF-7/ADR cells. Results EBB had antitumor effects for MCF-7 and MCF-7/ ADR. It could potentiate the antitumor effect of dororubicin with CDI of 0.73 and 0. 49 for MCF-7 and MCF- 7/ADR, respectively. EBB and doxorubicin acted synergistically in elevating apoptosis of MCF-7/ADR anddownregulating the expression of P-gp in a dose-dependent manner in MCF-7/ADR. EBB restored the intracellular accumulation of doxorubicin in McF-7/ADR cells in a dose-dependent manner. After pretreatment with EBB for 24 h and 48 h, the intracellular accumulation of doxorubicin and Rh123 was obviousely re- stored in MCF-7/ADR cells compared with control in a time-dependent manner. Conclusion EBB is a potential agent which has strong inhibitory effect on both multidrug resistant cells and their parental cells. EBB can significantly potentiate the antitumor effects of dororubicin in MCF-7/ADR cells by blocking the function of P-glycoprotein and inhibiting the expression of P-glycoprotein.

关 键 词：多药耐药 EBB MDR1基因 P-GP 阿霉素 共聚焦激光扫描显微镜 MCF-7/ADR细胞系 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学]