西妥昔联合阿霉素对三阴乳腺癌细胞增殖与凋亡的影响  

Effect of cetuximab combined with adriamycin on proliferation and apoptosis of triple negative breast cancer cells

作　　者：王秀[1] 李见春[1] 张竞竞[2] 赵素容[1] 刘浩[1] 

机构地区：[1]蚌埠医学院药学系,安徽蚌埠233030 [2]蚌埠医学院第一附属医院肿瘤内科,安徽蚌埠233003

出　　处：《中国药理学通报》2015年第12期1735-1740,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81372899);安徽省高等学校自然科学研究项目(No KJ2015B065by)

摘　　要：目的探讨西妥昔联合阿霉素对三阴乳腺癌细胞株MDA-MB-231增殖和凋亡的影响。方法 MTT法检测药物对细胞活性的影响;碘化丙啶(propidium iodide,PI)染色法测定药物对细胞凋亡率的影响;JC-1染色法测定线粒体膜电位的变化;Western blot测定细胞内葡萄糖调节蛋白78(glucose regulated protein 78,GRP-78)、Bcl-2及Caspase-3的表达水平。结果 MTT结果显示,西妥昔对MDA-MB-231细胞的抑制作用随浓度增加而增强,但作用时间对细胞的抑制作用影响较小。阿霉素对MDA-MB-231细胞的抑制作用具有浓度与时间依赖性。西妥昔与阿霉素合用能明显增强对MDA-MB-231细胞株的抑制作用,合用组12、24、48 h的细胞存活率与西妥昔组、阿霉素组相比,差异均具有显著性(P<0.05或P<0.01)。PI结果显示:单用西妥昔、阿霉素均可诱导MDA-MB-231的凋亡。联合用药可明显提高细胞的凋亡率,合用24 h的凋亡率达到了(43.86±3.62)%,与西妥昔组(11.0±2.32)%、阿霉素组(17.1±1.37)%相比,差异具有显著性(P<0.01)。JC-1染色结果显示西妥昔、阿霉素单用及联合应用均可降低线粒体膜电位,合用组降低更明显。Western blot显示西妥昔可下调Bcl-2、GRP-78的表达,激活Caspase-3。阿霉素对Bcl-2及Caspase-3表达无影响,可增加GRP-78的表达。合用组的GRP-78、Bcl-2表达明显降低,Caspase-3的激活明显增加。结论西妥昔与阿霉素合用可增强对三阴乳腺癌细胞株MDA-MB-231的抑制作用,增加细胞凋亡,其机制可能是西妥昔通过下调内质网应激水平,去除内质网应激对细胞的保护作用,进而减少Bcl-2表达,降低线粒体膜电位,激活线粒体凋亡通路,促进细胞的凋亡。Aim To detect the effects of cetuximab combined with adriamycin on the proliferation and apoptosis of triple-negative breast cancer cells. Methods Cell viability was evaluated by MTT assay. The cell apoptosis was analyzed by flow cytometry with propidium iodide staining. JC-1 staining was used to determine mitochondrial membrane potential. The expressions of glucose regulated protein78( GRP-78),Bcl-2and Caspase-3 were measured with Western blot. Results MTT assay showed that cetuximab had inhibition effect on the breast cancer cell MDA-MB-231 growth,and the effect was related to concentration of drug. The inhibition effect of adriamycin on MDA-MB-231 had remarkabe relationship with time and concentration. When combined with each other,they could remarkably increase inhibition effect. The viability of cells in combination group for 12 h,24 h,48 h,significantly lower than that in cetuximab or adriamycin group( P < 0. 05 or P < 0. 01). Apoptosis results showed that cell apoptosis was significantly increased when cetuximab combined with adriamycin,reached( 43. 93 ± 3. 59) % for 24 h,had remarkably statistical significance compared to cetuximab or adriamycin group( P < 0. 01). JC-1 staining indicated that cetuximab or adriamycin could reduce the mitochondrial membrane potential,but the reduction effect was more remarkable in the combination group. Western blot revealed that cetuximab could reduce the expression of GRP-78 and Bcl-2,and increased the expression of Caspase-3 and its activity. The expressions of Bcl-2,Caspase-3 had no significant change in adriamycin group,but GRP-78 was increased. In combination group,the expression of GRP-78 and Bcl-2 was significantly decreased,but Caspase-3 was increased notablely compared to adriamycin group. Conclusions The combination of cetuximab and adriamycin enhances the inhibition effect on the triple-negative breast cancer MDA-MB-231 cells,and increases cell apoptosis. The mechanism may be that cetuximab reduces the endoplasmic reticulum stress level,then activates the mitoc

关 键 词：三阴乳腺癌 联合用药 西妥昔 阿霉素 线粒体膜电位 凋亡 

分 类 号：R329.4[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]