紫杉醇耐药乳腺癌细胞外泌体来源的miR-5585-5p诱导乳腺癌细胞产生耐药表型研究  

Study on the drug-resistant phenotypes of breast cancer cells induced by exosomal miR-5585-5p derived from paclitaxel-resistant breast cancer cells

作　　者：魏欣宇 高鹏 刘佳欣 王文锐 杨清玲 陈昌杰 WEI Xin-yu;GAO Peng;LIU Jia-xin;WANG Wen-rui;YANG Qing-ling;CHEN Chang-jie(Key Laboratory of Cancer Translational Medicine of Anhui Province,Bengbu Anhui 233000,Bengbu Anhui 233000;School of Clinical Medicine,Bengbu Medical College,Bengbu Anhui 233030,China)

机构地区：[1]癌症转化医学安徽省重点实验室,安徽蚌埠233000 [2]蚌埠医学院临床医学院,安徽蚌埠233030

出　　处：《蚌埠医学院学报》2021年第8期989-993,共5页Journal of Bengbu Medical College

基　　金：安徽省高校自然科学研究重大项目(KJ2019ZD28);蚌埠医学院科技计划重点项目(BYKY2019265ZD);国家级大学生创新创业训练计划项目(20210367032);蚌埠医学院研究生科研项目(Byycx 20019)。

摘　　要：目的:研究紫杉醇耐药乳腺癌细胞外泌体来源的miR-5585-5p对乳腺癌细胞迁移、凋亡和耐药性的影响。方法:超速离心法获取外泌体,使用qRT-PCR方法分别检测乳腺癌细胞(SKBR-3)和紫杉醇耐药乳腺癌细胞(SKBR-3/PR)细胞株和外泌体中miR-5585-5p的表达量;通过外泌体共培养技术以及在乳腺癌耐药细胞中转染miR-5585-5p的抑制剂,用Transwell检测其迁移能力,流式细胞术检测其凋亡情况;qRT-PCR和Western blotting检测其耐药指标。结果:与SKBR-3相比,miR-5585-5p在耐药细胞SKBR-3/PR细胞株和外泌体中表达均上调(P<0.01和P<0.05);在耐药细胞株中转染miR-5585-5p的抑制剂,Transwell、流式分析等结果显示,与对照相比,抑制剂组的耐药性下降,生物学活性降低(P<0.01);耐药细胞源性的外泌体与亲本细胞共培养可使SKBR-3细胞产生耐药表型,增强其迁移能力,抑制其凋亡率(P<0.01),而转染抑制剂的耐药细胞分泌的外泌体可减弱这一作用。结论:紫杉醇耐药乳腺癌细胞通过外泌体递送高表达的miR-5585-5p至亲本细胞并诱导亲本细胞产生耐药表型,外泌体miR-5585-5p可为乳腺癌耐药治疗提供新的分子靶点,为乳腺癌预后评估提供新的生物标志物。Objective:To study the effects of exosomal miR-5585-5p derived from paclitaxel-resistant breast cancer cells on the migration,apoptosis and drug resistance of breast cancer cells.Methods:The exosomes were obtained by ultracentrifugation,and the expression levels of miR-5585-5p in breast cancer cell line(SKBR-3),paclitaxel-resistant breast cancer cell line(SKBR-3/PR)and exosomes were detected using qRT-PCR.The inhibitor of miR-5585-5p was transfected into paclitaxel-resistant breast cancer cells,the migration ability of cells was detected using Transwell assay,and the apoptosis of cells was detected using flow cytometry.The qRT-PCR and western blotting were used to detect the expression levels of drug-resistant gene.Results:Compared with SKBR-3 cell line,the expression levels of miR-5585-5p in drug-resistant SKBR-3/PR cell line and exosomes were up-regulated(P<0.01 and P<0.05).After the miR-5585-5p inhibitor was transfected into the drug-resistant cell line,the results of Transwell assay and flow cytometry showed that the drug resistance and biological activity in the inhibitor group decreased compared with the control group(P<0.01).The co-cultured drug-resistant cell-derived exosomes with breast cancer cells could induce the drug-resistant phenotype,enhance the ability of migration and inhibit the apoptosis rate of SKBR-3 cells(P<0.01).However,the exosomes derived from drug-resistant breast cancer cells transfected with miR-5585-5p inhibitor could reduce the effects.Conclusions:The paclitaxel-resistant breast cancer cells can deliver the highly expressed miR-5585-5p to the parent cells through exosomes,and induce the parent cells to produce drug-resistant phenotypes.The exosome miR-5585-5p can provide new molecular targets for the treatment of breast cancer drug-resistance,and provide new biomarkers for the prognosis assessment of breast cancer.

关 键 词：乳腺肿瘤 紫杉醇耐药 外泌体 miR-5585-5p 

分 类 号：R737.9[医药卫生—肿瘤]