益气小复方抑制三阴性乳腺癌外泌体传递耐药信息的作用机制研究  

Mechanisms of Yiqi Compound Recipe on inhibiting exosomes transmitting drug resistance information in triple negative breast cancer cells

作　　者：王冰[1] 张玉柱[1] 陈红风[1] WANG Bing;ZHANG Yuzhu;CHEN Hongfeng(Breast Department of Chinese Medicine,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院中医乳腺科,上海200032

出　　处：《上海中医药大学学报》2018年第6期50-55,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81373647)

摘　　要：目的:探讨三阴性乳腺癌细胞(MDA-MB-231)顺铂(DDP)耐药细胞株释放的外泌体(Exosomes,EXOs)在细胞间可能存在的耐药信息传递作用及益气小复方对外泌体传递耐药信息的阻滞作用。方法:以三阴性乳腺癌亲本细胞MDA-MB-231及其DDP耐药细胞(MDA-MB-231/DDP)为模型,采用过滤离心法提取MDA-MB-231/DDP外泌体(DDP-EXO);透射电子显微镜观察鉴定细胞DDP-EXO形态; Western blot检测其CD63、TSG101、CD9的表达;用激光共聚焦显微镜追踪DDP-EXO与敏感细胞MDA-MB-231的相互作用; MTT法检测MDA-MB-231与DDP-EXO共培养后对DDP的IC50的影响及益气小复方干预DDP-EXO与MDA-MB-231细胞共培养后对DDP IC50的影响。结果:透射电子显微镜下观察DDP-EXO呈现典型的圆形或椭圆形杯口状结构,直径40~100 nm; DDP-EXO表达外泌体标志蛋白CD63、TSG101、CD9;用PKH67标记DDP-EXO显示,DDP-EXO可以被敏感细胞MDA-MB-231摄取;敏感细胞株MDA-MB-231与DDP-EXO共培养48 h后,DDP的IC50升高2.24倍,差异有统计学意义(P<0.01); MDA-MB-231细胞与DDP-EXO共培养,再予益气小复方干预后DDP IC50明显降低(P<0.01)。结论:三阴性乳腺癌外泌体可能具有传递耐药信息的作用,耐药细胞外泌体能够使敏感细胞获得一定耐药性,益气小复方可一定程度上阻滞外泌体传递耐药性。Objective: To study the potential function of exosomes ( EXOs) released by triple negative breast cancer platinum( DDP) resistant cells ( MDA-MB-231 /DDP) in transmitting drug resistance information between cells,and the inhibitory effects of YiqiCompound Recipe on exosomes transmitting drug resistance information. Methods: MDA-MB-231 as the parent cells of triple negativebreast cancer and its resistant cell MDA-MB-231 /DDP were used as model. The filtration centrifugation method was used to extract theexosomes of MDA-MB-231 /DDP ( DDP-EXO) . The morphology of DDP-EXO was observed and identified by transmission electronmicroscope. The expressions of CD63,TSG101 and CD9 were detected by Western blot. The interaction between DDP-EXO and sensitivecells MDA-MB-231 was traced by confocal laser scanning microscope. Before and after treatment with Yiqi Compound Recipe,the effectof co-culture of DDP-EXO and MDA-MB-231 on IC50 of DDP was detected by MTT assay. Results: Under the transmission electronmicroscope,DDP-EXO presented a typical round or oval cup-like structure with a diameter of 40-100 nm. The marker proteins ofexosomes including CD63,TSG101 and CD9 were expressed in DDP-EXO. DDP-EXO labeled with PKH67 could be up-taken by sensitiveMDA-MB-231 cells. After co-culture with DDP-EXO for 48 hours,the IC50 value of MDA-MB-231 to DDP was increased by 2.24 times,with statistically significant difference( P<0.01) . After treatment with Yiqi Compound Recipe on co-culture of DDP-EXO and MDA-MB-231,the IC50 value of MDA-MB-231 to DDP was obviously decreased ( P>0.01) . Conclusion: Triple negative breast cancer exosomes mayplay the role of transmitting resistance information. Drug resistant cell exosomes enables sensitive cells to acquire certain drug resistance.Yiqi Compound Recipe can inhibit exosomes transmitting drug resistance information to a certain extent.

关 键 词：三阴性乳腺癌 益气小复方 外泌体 耐药 MDA-MB-231细胞 顺铂 

分 类 号：R285[医药卫生—中药学]