细胞外囊泡介导的微小RNA-491递送抑制骨肉瘤肺转移  

Extracellular vesicle-mediated delivery of microRNA-491 inhibits lung metastasis of osteosarcoma

作　　者：刘曌 高祥彬 赵锡江[1] Liu Zhao;Gao Xiangbin;Zhao Xijiang(Department of Orthopedics,Affiliated Hospital of Jiangnan University,Wuxi 214062,China)

机构地区：[1]江南大学附属医院骨科,无锡214062

出　　处：《中华实验外科杂志》2024年第1期120-123,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨细胞外囊泡(EV)介导的微小RNA(miR)-491递送抑制骨肉瘤肺转移。方法细胞水平检测miR-491靶向降解胫骨来源的骨肉瘤细胞中αB-晶状体蛋白(CRYAB)的信使RNA(mRNA)和蛋白,分为6组,miR-NC组、miR-491组、Vector+miR-NC组、Vector+miR-491组、CRYAB+miR-NC组、CRYAB+miR-491组。收集我院接受骨折治疗手术的男性患者皮下脂肪组织并分离脂肪组织来源的间充质间质细胞(AD-MSC),在AD-MSC中过表达miR-491或阴性对照(NC)后收集细胞外囊泡miR-491-EV和NC-EV。随后建立小鼠骨肉瘤肺转移模型并分为两组:NC-EV组和miR-491-EV组,检测肺转移结节数和肺重量。通过t检验和方差分析检测不同组间差异。结果miR-491组的CRYAB的mRNA和蛋白水平低于miR-NC组(1.00±0.03比0.19±0.02,t=38.912,P<0.01)。Vector+miR-491组的侵袭能力和迁移能力低于Vector+miR-NC组(1.00±0.08比0.31±0.03,t=31.284,P<0.01;1.00±0.08比0.28±0.03,t=32.641,P<0.01);CRYAB+miR-NC组的侵袭能力和迁移能力高于Vector+miR-NC组(1.00±0.08比3.22±0.18,t=43.650,P<0.01;1.00±0.08比1.88±0.23,t=14.000,P<0.01)。miR-491-EV组的鼠骨肉瘤转移性肺结节的数量(6.03±1.21比1.02±0.23,F=154.412,P<0.05)和肺重量(0.68±0.09比0.47±0.04,F=43.254,P<0.05)低于NC-EV组。结论miR-491能够通过下调CRYAB在骨肉瘤中的表达作为肿瘤抑制因子。Objective To investigate the inhibitory effect of extracellular vesicle(EV)-mediated microRNA(miR)-491 delivery on pulmonary metastasis of osteosarcoma.Methods MiR-491 was used to target the degradation ofαB-crystallin(CRYAB)mRNA and protein in tibial origin osteosarcoma cells at the cellular level,and the following groups were set up:miR-NC group,miR-491 group,vector+miR-NC group,vector+miR-491 group,CRYAB+miR-NC group,CRYAB+miR-491 group.Subcutaneous adipose tissue of male patients undergoing fracture surgery in our hospital was collected and adipose-derived mesenchymal interstitium cells(AD-MSCs)were isolated.Extracellular vesicles Mir-491-EV and NC-EV were collected after miR-491 was overexpressed in AD-MSCs or negative control(NC).Subsequently,the mouse osteosarcoma lung metastasis model was established and divided into two groups:NC-EV group and miR-491-EV group,and the number of lung metastasis nodules and lung weight were detected.The difference between different groups was detected by t test and ANOVA.Results CRYAB mRNA and protein levels in miR-491 group were lower than those in miR-NC group(1.00±0.03 vs.0.19±0.02,t=38.912,P<0.01).The invasion and migration ability of vector+miR-491 group was lower than that of vector+miR-NC group(1.00±0.08 vs.0.31±0.03,t=31.284,P<0.01;1.00±0.08 vs.0.28±0.03,t=32.641,P<0.01).The invasion and migration ability of CRYAB+miR-NC group was higher than that of vector+miR-NC group(1.00±0.08 vs.3.22±0.18,t=43.650,P<0.01;1.00±0.08 vs.1.88±0.23,t=14.000,P<0.01).The number of metastatic pulmonary nodules in miR-491-EV group(6.03±1.21 vs.1.02±0.23,F=154.412,P<0.05)and lung weight(0.68±0.09 vs.0.47±0.04,F=43.254,P<0.05)were lower than those in NC-EV group.Conclusion MiR-491 can act as a tumor suppressor by down-regulating CRYAB expression in osteosarcoma.miR-491 enriched EV derived from AD-MSCs can be used as a potential therapy for metastatic osteosarcoma.

关 键 词：细胞外囊泡 微小RNA-491 骨肉瘤 肺转移 αB-晶状体蛋白 

分 类 号：R738.1[医药卫生—肿瘤]