乳腺癌耐阿霉素细胞MCF-7/ADR外泌体传递tTG参与MCF-7细胞耐药的研究  

Exosomes of breast cancer adriamycin-resistant MCF-7/ADR cells transfer tTG to participate in drug resistance of MCF-7 cells

作　　者：张桓海 沈媛媛 王晓红[2] 张湘生 程凯 ZHANG Huanhai;SHEN Yuanyuan;WANG Xiaohong;ZHANG Xiangsheng;CHENG Kai(Yantai Affiliated Hospital of Binzhou Medical University,Shandong Yantai 264100,China;Binzhou Medical University Hospital,Shandong Binzhou 256600,China.)

机构地区：[1]滨州医学院烟台附属医院,山东烟台264100 [2]滨州医学院附属医院,山东滨州256600

出　　处：《现代肿瘤医学》2024年第3期393-397,共5页Journal of Modern Oncology

基　　金：山东省医药卫生科技发展计划项目(编号:202104010967);滨州医学院“临床+X”项目(编号:滨医行发[2022]22号)。

摘　　要：目的:研究乳腺癌耐阿霉素细胞MCF-7/ADR通过外泌体传递tTG参与敏感细胞MCF-7耐药的过程。方法:本实验首先培养MCF-7、MCF-7/ADR细胞,然后检测MCF-7、MCF-7/ADR细胞中外泌体的含量,并将外泌体提取后备用。MCF-7/ADR细胞外泌体(EXO/ADR)与MCF-7细胞共培养建立MCF-7的外泌体耐药细胞(MCF-7+EXO/ADR)。使用Western blot法检测MCF-7、MCF-7/ADR、MCF-7+EXO/ADR细胞中的tTG蛋白的表达水平,使用流式细胞术分别检测敏感细胞株外泌体(EXO/S)、耐药细胞株外泌体(EXO/ADR)中tTG的表达水平。使用CCK-8法分别检测MCF-7、MCF-7/ADR、MCF-7+EXO/ADR细胞对阿霉素的敏感性情况。结果:实验结果显示,在EXO/S、EXO/ADR中均可以表达CD63、TSG101,但是较低表达Calnexin。经过流式细胞术检测发现,EXO/S、EXO/ADR中的tTG表达水平分别为(18.3±3.63)%、(46.07±8.31)%,二者tTG的表达有明显差异。经过Western blot检测MCF-7、MCF-7/ADR、MCF-7+EXO/ADR细胞中tTG的表达情况,我们发现MCF-7+EXO/ADR细胞的tTG表达水平较MCF-7细胞升高。MCF-7、MCF-7/ADR和MCF-7+EXO/ADR细胞对阿霉素的IC50分别为(1.91±0.18)μg/mL、(82.11±3.51)μg/mL、(6.16±1.15)μg/mL。结论:MCF-7/ADR细胞可能经分泌外泌体来传递tTG参与MCF-7细胞对阿霉素耐药的过程。Objective:To study the process of breast cancer adriamycin-resistant cell MCF-7/ADR participating in the drug resistance of sensitive cell MCF-7 through tTG transmitted by exosomes.Methods:This experiment first cultured MCF-7 and MCF-7/ADR cells,and then measured the content of exosomes in MCF-7 and MCF-7/ADR cells,and extracted the exosomes for later use.MCF-7/ADR exosomes(EXO/ADR)were co cultured with MCF-7 cells to establish MCF-7 exosomes resistant cells(MCF-7+EXO/ADR).Western blot was used to detect the expression level of tTG protein in MCF-7,MCF-7/ADR,and MCF-7+EXO/ADR cells.Flow cytometry was used to detect the expression level of tTG protein in sensitive cell lines exosomes(EXO/S)and drug-resistant cell lines exosomes(EXO/ADR),respectively.CCK-8 method was used to detect the drug sensitivity of MCF-7,MCF-7/ADR,and MCF-7+EXO/ADR to doxorubicin.Results:Both EXO/S and EXO/ADR expressed CD63 and TSG101,but low expression of Calnexin.Flow cytometry results showed that the tTG expression level of EXO/S was(18.3±3.63)%,and that of EXO/ADR was(46.07±8.31)%.The expression level of tTG protein in EXO/ADR was significantly higher than that in EXO/S,and the difference was statistically significant(P<0.05).Western blot was used to detect the expression level of tTG in MCF-7,MCF-7/ADR and MCF-7+EXO/ADR groups,and the tTG expression level of MCF-7+EXO/ADR increased.The IC50 of MCF-7/ADR cells and MCF-7+EXO/ADR cells against adriamycin was(1.91±0.18)μg/mL,(82.11±3.51)μg/mL,and(6.16±1.15)μg/mL.Conclusion:MCF-7/ADR cells may secrete exosomes to transfer tTG and participate in the process of adriamycin resistance of MCF-7.

关 键 词：外泌体 TTG 乳腺癌 阿霉素耐药 

分 类 号：R737.9[医药卫生—肿瘤]