肿瘤相关巨噬细胞来源外泌体通过miR-21促进前列腺癌细胞紫杉醇耐药的机制研究  

Mechanism of tumor-associated macrophage-derived exosomes promoting paclitaxel resistance of prostate cancer cells via miR-21

作　　者：梁永钢 肖华 董海波 谢林森 LIANG Yonggang;XIAO Hua;DONG Haibo;XIE Linsen(Department of Laboratory Medicine,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Henan Zhengzhou 450001,China;Department of Laboratory Medicine,Zhengzhou Yihe Hospital,Henan Zhengzhou 450018,China)

机构地区：[1]郑州大学附属郑州中心医院检验科,河南郑州450001 [2]郑州颐和医院检验科,河南郑州450018

出　　处：《现代肿瘤医学》2023年第15期2782-2788,共7页Journal of Modern Oncology

基　　金：河南省医学科技攻关计划联合共建项目(编号:LHGJ20200778)。

摘　　要：目的:探究外泌体(Exos)对miR-21的调控作用及对前列腺癌(PC)细胞紫杉醇耐药的影响。方法:取前列腺癌组织及癌旁组织,免疫组化法检测CD68、M1型巨噬细胞表面标记(CD86)、M2型巨噬细胞表面标记(CD206)阳性表达,以鉴定巨噬细胞浸润及表型变化;取人巨噬细胞株THP-1诱导建立M2型巨噬细胞极化模型,差速离心法提取Exos,与前列腺癌细胞PC-3共培养,设置为:PC-3组、PC-3+Exos组、PC-3+多西紫杉醇(DTX)组、PC-3+Exos+DTX组;CCK-8法检测细胞增殖活性;Transwell检测细胞迁移能力;qRT-PCR法检测Exos及细胞中miR-21表达;Western blot法检测细胞耐药蛋白(P-gp、Txr1)水平。敲低Exos中miR-21表达或阻断巨噬细胞Exos分泌后,与PC-3共培养,并植入裸鼠皮下,给予DTX干预治疗,测量瘤体体积及瘤体重量,以验证Exos对miR-21调控及PC-3耐药性的影响。结果:PC癌组织中主要以M2型巨噬细胞为主。M2型Exos直径约为100 nm,呈杯状粒子形状,可被PC-3细胞摄取,其高表达miR-21。与PC-3组相比,M2型Exos可促进PC-3细胞增殖、迁移,抑制凋亡,促进PC-3细胞高表达miR-21及耐药蛋白P-gp、Txr1(P<0.05)。DTX干预治疗下,M2型Exos也可进一步促进PC-3细胞增殖、迁移,加剧PC对DTX药物的耐受性(P<0.05)。敲低Exos中miR-21表达或阻断Exos分泌,均可减小瘤体体积及重量,增加DTX化疗的敏感性(P<0.05)。结论:M2型巨噬细胞Exos可通过传递miR-21,促进PC细胞增殖、迁移,加剧PC对DTX药物的耐受性。Objective:To explore the regulatory effect of exosomes(Exos)on miR-21 and its effect on paclitaxel resistance in prostate cancer(PC)cells.Methods:Prostate cancer tissues and adjacent tissues were collected,and the positive expression of CD68,M1 macrophage surface marker(CD86),and M2 macrophage surface marker(CD206)was detected by immunohistochemistry,and then macrophage infiltration and phenotypic changes were identified.The human macrophage cell line THP-1 was used to induce the establishment of M2 macrophage polarization model,and Exos was extracted by differential centrifugation,and co-cultured with prostate cancer cells PC-3,which were set up PC-3 group,PC-3+Exos group,PC-3+Docetaxel(DTX)group,and PC-3+Exos+DTX group.Cell proliferation activity was detected by CCK-8.Cell migration ability was detected by Transwell.The expression of miR-21 in Exos and cells were detected by qRT-PCR.The levels of cell resistance proteins(P-gp,Txr1)were detected by Western blot.After knockdown the expression of miR-21 in Exos or blocking the secretion of Exos in macrophages,it was co-cultured with PC-3 and implanted subcutaneously in nude mice.After DTX intervention,tumor volume and weight were measured to verify the influences of Exos on miR-21 regulation and PC-3 drug resistance.Results:M2 macrophages were the predominant cells in PC cancer tissues.M2-type Exos had a diameter of about 100 nm and a cup-like particle shape.They were able to be taken up by PC-3 cells,and highly expressed miR-21.Compared with PC-3 group,M2-type Exos were was able to promote PC-3 cell proliferation,migration,inhibit apoptosis and promote the expression of miR-21 and drug-resistant proteins P-gp and Txr1(P<0.05).Under DTX intervention,M2-type Exos were able to further promote the proliferation and migration of PC-3 cells,and aggravate the resistance of PC to DTX(P<0.05).Knockdown the expression of miR-21 in Exos or blocking the secretion of Exos was able to reduce tumor volume and weight,and increase the sensitivity to DTX chemotherapy(P<0.05).Concl

关 键 词：肿瘤相关巨噬细胞 外泌体 前列腺癌 耐药 紫杉醇 微小RNA-21 

分 类 号：R737.25[医药卫生—肿瘤]