宫颈癌细胞分泌外泌体介导上皮-间质转化提高癌前细胞侵袭能力的体外研究  

Exosomes derived from cervical carcinoma cells promote preneoplastic cell invasion in vitro via epithelial mesenchymal transition

作　　者：易红艳[1] 周琛斐 梁莉[2] 王薇[1] 魏文斐[1] 陈晓静[1] 吴湘光 严瑞明 刘国炳[1] 陈春林[1] 钟梅[1] 余艳红[1] Yi Hongyan Zhou Chenfei Liang Li et al(Department of Obstetrics and Gynecology Department of Pathology, Nanfang Hospital, South- ern Medical University, Guangzhou 510515)

机构地区：[1]南方医科大学南方医院妇产科,广州510515 [2]南方医科大学南方医院病理科,广州510515

出　　处：《现代妇产科进展》2017年第3期165-168,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金(No:81072132;81372871;81672589;JCYJ20160429161218745)

摘　　要：目的:研究宫颈癌细胞Siha分泌的外泌体(Exosomes)能否介导上皮-间质转化(EMT),从而改变癌前细胞Ect1的体外侵袭能力。方法:差速超速离心法分离SihaExo,透射电镜观察形态,Western blot检测标志蛋白表达,共聚焦显微镜观察PKH67染色外泌体的生物学活性。Siha-Exo和PBS分别与Ect1共培养48h后Western blot检测上皮和间质转化标志蛋白的表达,Transwell侵袭实验检测共培养后Ect1体外侵袭能力。结果:Siha-Exo为直径30~100nm的盘状囊泡,特异性表达标志性蛋白CD63和CD81,能够被受体细胞Ect1内吞,具备生物学活性。Western blot结果显示,与PBS组相比,Siha-Exo组下调Ect1细胞中上皮标志蛋白E-Cadherin和β-Catenin的表达,上调间质标志蛋白NCadherin和Vimentin的表达。Transwell侵袭实验显示,与PBS组相比,Siha-Exo组穿透聚碳酯膜的Ect1细胞数明显增多(P=0.01)。结论:Siha-Exo能够介导Ect1发生EMT变化,提高其体外侵袭能力。Objective： To investigate whether Exosomes secreted by cervical cancer cell lines Siha（ Siha-Exo） have ability to promote Ect1 invasion via epithelial mesenchymal transition（ EMT） in cervical cancer. Methods： Morphology and marker protein of Siha-Exo isolated by different ultracentrifugation were identified by transmission electron microscopy and Western blot.And green fluorescent PKH67-labeled Siha-Exo detected by confocal images for biological activity.Ect1 was co-cultured with Siha-Exo or PBS for 48 hours. The expression of epithelial and mesenchyme markers were examined by Western blot. Transwell chambers were used in vitro invasion assay.Results： Siha-Exo exhibited typical cup-shaped vesicles and a size range of30-100 nm,expressed exosomal marker protein CD63 and CD81,and were biologically uptaked by recipient Ect1. Compared with PBS treated cells,Ect1 treated with Siha-Exo decreased expression of epithelial marker E-Cadherin and β-Catenin,and increased expression of mesenchymal marker N-Cadherin and Vimentin.Meanwhile in Siha-Exo group,numbers of Ect1 penetrating transwell membrane significantly increased in contrast with PBS group（ P = 0.01）.Conclusion： Siha-Exo elicited Ect1 to an EMT-like phenotype,thus promoting invasion in vitro.

关 键 词：外泌体 宫颈癌 上皮间质转化 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]