外泌体中circ-RPPH1通过circ-RPPH1/miR-146b-3p/MUC19通路轴调控乳腺癌对紫杉醇的耐药性  

Exosomes circ-RPPH1 regulates breast cancer paclitaxel resistance through the circ-RPPH1/miR-146b-3p/MUC19 axis

作　　者：赵玥 付永强 周真林 杨明 黄维超 方雅兰 ZHAO Yue;FU Yongqiang;ZHOU Zhenlin;YANG Ming;HUANG Weichao;FANG Yalan(Department of Thyroid Breast Hernia Surgery, the Affiliated Guizhou Aviation Industry Cor Ltd No 300 Hospital of Zunyi Medical University, Guiyang 550025, China;Zhuhai Maternal and Child Health Care Hospital, Zhuhai 519000, China)

机构地区：[1]遵义医科大学附属贵航三00医院甲乳疝外科,贵州贵阳550025 [2]珠海市妇幼保健院病理科,广东珠海519000

出　　处：《中国肿瘤外科杂志》2022年第3期296-306,共11页Chinese Journal of Surgical Oncology

基　　金：贵州省卫健委科学技术基金项目(gzwjkj2020-1-094)。

摘　　要：目的 研究外泌体circ-RPPH1在乳腺癌患者血清、乳腺癌细胞系MCF-7、MDA-MB-231及乳腺癌紫杉醇耐药细胞系MCF-7/PTX、MDA-MB-231/PTX中的表达情况,探讨circ-RPPH对乳腺癌耐药细胞株生物学功能的影响及作用机制。方法 通过生物信息数据检索系统分析乳腺癌组织与正常组织差异化表达circ-RPPH1及其下游靶向miR-146b-3p、MUC19通路;收集2020年2月至2021年10月遵义医科大学附属贵航三OO医院诊治的53例乳腺癌患者,分为紫杉醇敏感组(24例)与紫杉醇耐药组(29例)。取两组患者血清中外泌体,提取乳腺癌细胞系MCF-7、MDA-MB-231及乳腺癌紫杉醇耐药细胞系MCF-7/PTX、MDA-MB-231/PTX中的外泌体,检测circ-RPPH1的表达水平;通过外泌体摄取实验验证外泌体参与介导circ-RPPH1的转运;siRNA转染细胞后观察circ-RPPH1表达与乳腺癌细胞耐药、增殖、周期、迁移和侵袭情况相关性;构建稳转circ-RPPH1裸鼠移植瘤模型,观察circ-RPPH1水平与瘤体生长的关系并测定瘤体中circ-RPPH1含量。结果 circ-RPPH1在乳腺癌组织中差异化表达;circ-RPPH1在紫杉醇耐药患者血清及乳腺癌紫杉醇耐药细胞系MCF-7/PTX、MDA-MB-231/PTX中表达升高(P<0.001);与紫杉醇敏感组比较,紫杉醇耐药组血清外泌体中circ-RPPH1表达升高(P<0.001);外泌体负责转运circ-RPPH1;摄取紫杉醇耐药细胞外泌体后乳腺癌细胞系circ-RPPH1表达增高;敲低circ-RPPH1的表达后乳腺癌细胞系及乳腺癌耐药细胞系增殖、迁移及侵袭能力降低,细胞合成受阻;双荧光素实验及RNA免疫共沉淀表明,circ-RPPH1/miR-146b-3p/MUC19之间存在靶向调控关系;与紫杉醇敏感组比较,紫杉醇耐药组血清外泌体中miR-146b-3p表达降低,MUC19表达升高(均P<0.001);敲低circ-RPPH1可以抑制肿瘤的生长,且肿瘤中circ-RPPH1含量减少。结论 外泌体中circ-RPPH1通过circ-RPPH1/miR-146b-3p/MUC19通路轴增加乳腺癌对紫杉醇的耐药性,影响乳腺癌的恶性生物学行Objective To investigate the expression of exosome circ-RPPH1 in serum of breast cancer patients,breast cancer cell lines MCF-7,MDA-MB-231 and breast cancer paclitaxel-resistant cell lines MCF-7/PTX,MDA-MB-231/PTX,revealing the effect and mechanism of circ-RPPH1 on the biological function of drug-resistant breast cancer cell lines.Methods The differential expression of circ-RPPH1 and its downstream targeting miR-146b-3p and MUC19 pathways in breast cancer tissues and normal tissues were analyzed through the bioinformatics data retrieval system.A total of 53 breast cancer patients diagnosed and treated in the Affiliated Guizhou Aviation Industry Cor Ltd No 300 Hospital of Zunyi Medical University from February 2020 to October 2021 were collected and divided into a taxane-sensitive group(24 cases)and a paclitaxel-resistant group(29 cases).To extract exosomes from breast cancer cell lines MCF-7,MDA-MB-231 and breast cancer paclitaxel-resistant cell lines MCF-7/PTX,MDA-MB-231/PTX,and assess the expression of circ-RPPH1.Exosome uptake experiments verifies that exosomes are involved in mediating the transport of circ-RPPH1.While the expression levels of circ-RPPH1 were lower,the correlation of circ-RPPH1 expression with breast cancer drug resistance,proliferation,clone formation,cycle,migration and invasion was observed.A nude mouse xenograft model of stably transfected circ-RPPH1 was constructed to observe the relationship between the level of circ-RPPH1 and tumor growth and to determine the content of circ-RPPH1 in the tumor.Results circ-RPPH1 was differentially expressed in breast cancer tissues.The expression of circ-RPPH1 was increased in the serum of paclitaxel-resistant patients and in the paclitaxel-resistant breast cancer cell lines MCF-7/PTX and MDA-MB-231/PTX(P<0.001).Compared with paclitaxel sensitive group,the expression of circ-RPPH1 in the serum exosomes of paclitaxel-resistant breast cancer patients was increased(P<0.001).Exosomes were responsible for transporting circ-RPPH1.While the expression levels

关 键 词：乳腺癌 外泌体 circ-RPPH1 化疗耐药 

分 类 号：R737.9[医药卫生—肿瘤]