MK-2206逆转人乳腺癌细胞耐药的作用及机制研究  

Reversal effect of MK-2206 on drug resistance in breast cancer cells and its mechanism

作　　者：焦明文 袁凯[1] 王玉龙[1] 陈鸿强[1] 周婷[1] 付荣湛[1] 

机构地区：[1]山东大学附属千佛山医院两腺外科,山东济南250014

出　　处：《中国普通外科杂志》2014年第11期1494-1500,共7页China Journal of General Surgery

基　　金：山东省自然科学基金资助项目(ZR2013HM040)

摘　　要：目的:探讨PI3K/Akt信号通路抑制剂MK-2206对人乳腺癌细胞耐药的逆转作用及机制。方法:用CCK-8法检测阿霉素与MK-2206对MCF-7细胞(阿霉素敏感)与MCF-7/ADR(阿霉素耐药)细胞生长的抑制情况并计算各自IC50值。根据IC50结果,选择低毒浓度阿霉素单独或联合不同浓度MK-2206处理MCF-7细胞和MCF-7/ADR细胞后,分别用CCK-8法和流式细胞术检测MK-2206对阿霉素耐药与阿霉素诱导细胞凋亡的影响,以及对MCF-7/ADR细胞内阿霉素蓄积的影响。用MK-2206单独作用MCF-7细胞与MCF-7/ADR细胞后,采用Western blot法检测PI3K/Akt信号通路相关蛋白的表达。结果:阿霉素与MK-2206作用后,两种细胞的增殖均受到明显抑制,阿霉素对MCF-7/ADR的IC50值明显高于MCF-7的IC50值(P<0.05),而MK-2206对两种细胞的IC50差异无统计学意义(P>0.05)。联合MK-2206可明显降低阿霉素对两种细胞IC50值,但MCF-7/ADR细胞的降低程度明显大于MCF-7细胞(P<0.05);MK-2206对阿霉素诱导的MCF-7细胞凋亡影响不明显,但能明显增加MCF-7/ADR细胞的凋亡率(P<0.05)。不同浓度MK-2206对MCF-7/ADR细胞内阿霉素的蓄积差异无统计学意义(P>0.05)。单独MK-2206处理后,MCF-7细胞p-(Thr308)Akt蛋白表达明显下调(P<0.01),但p-(Thr246)PRAS40蛋白表达无明显改变(P>0.05);MCF-7/ADR细胞以上两种蛋白的表达均明显下调(均P<0.05)。结论:MK-2206可以通过抑制PI3K/Akt信号通路来部分逆转人乳腺癌细胞的耐药性,且乳腺癌细胞耐药可能主要与该通路的PRAS40蛋白活化有关。Objective： To investigate the reversal effect of the PI3K/Akt singling pathway inhibitor MK-2206 on drug resistance in human breast cancer cells and its mechanism.Methods： The inhibitory effects of doxorubicin and MK-2206 on growth of MCF-7 cells（doxorubicinsensitive） and MCF-7/ADR cells（doxorubicin-resistant） were examined by CCK-8 assay and their IC50 values were calculated. Based on the IC50 results, MCF-7 and MCF-7/ADR cells were treated with a selected lowtoxic concentration of doxorubicin alone or combined with different concentrations of MK-2206, and then the inl uence of MK-2206 on doxorubicin-resistance and doxorubicin-induced apoptosis, and on doxorubicin accumulation in MCF-7/ADR cells were detected by CCK-8 assay and l ow cytometry, respectively. MCF-7 and MCF-7/ADR cells were treated with MK-2206 alone, and then the expressions of the proteins associated with PI3K/Akt signaling pathway were determined by Western blot.Results： h e proliferations in both types of cells were signii cantly inhibited by either doxorubicin or MK-2206 treatment, and the IC50 value of doxorubicin for MCF-7/ADR cells was signii cantly higher than that for MCF-7 cells（P〈0.05）, while the IC50 values between MK-2206 for the both type of cells had no signii cant dif erence（P〉0.05）. h e IC50 values of doxorubicin for both types of cells were decreased by combination treatment of MK-2206, but the decreasing degree in MCF-7/ADR cells was signii cantly greater than that in MCF-7 cells（P〈0.05）. MK-2206 exerted no obvious inl uence on the doxorubicin-induced apoptosis in MCF-7 cells, but signii cantly increased the doxorubicin-induced apoptosis in MCF-7/ADR cells（P〈0.05）. No signii cant dif erence was noted in doxorubicin accumulation among MCF-7/ADR cells treated with various concentrations of MK-2206（P〉0.05）. At er MK-2206 treatment alone, the protein expression of p-（h r308） Akt was signii cantly down-regulated（P〈0.05）, and the protein expression of p-（h r246） PA S40 showed n

关 键 词：乳腺肿瘤 抗药性 肿瘤 1-磷脂酰肌醇3-激酶 

分 类 号：R737.9[医药卫生—肿瘤]