摘要
目的:探讨癌细胞来源的外泌体携miR-205在鼻咽癌进展中的作用。方法:采用试剂盒提取法提取SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1鼻咽癌细胞系及正常鼻咽上皮细胞NP69中的外泌体,采用电镜检测外泌体形态,采用Western blot法检测外泌体特异性标志物CD63及Flotillin蛋白表达,采用实时荧光定量PCR法检测外泌体中miR-205的表达。结果:从SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1鼻咽癌细胞系中分离出椭圆状或圆形的泡状物,其中SUNE-1-5-8F细胞来源泡状物大小不均,直径40~130 nm,有完整的包膜;SUNE-1-6-10B细胞来源泡状物大小较为均一,直径约80 nm,有完整的包膜,且泡状物内电子致密物多;SUNE-1细胞来源泡状物大小均一,直径约100 nm,有完整的双层包膜,包膜光滑清晰,且泡状物内电子致密物较多。来源于SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1鼻咽癌细胞外泌体中的CD63及Flotillin蛋白表达量均显著高于来源于正常鼻咽上皮细胞NP69(P<0.05)。来源于SUNE-1鼻咽癌细胞外泌体中的miR-205表达量显著高于来源于正常鼻咽上皮细胞NP69(P<0.05)。结论:从鼻咽癌细胞中提取并鉴定获得外泌体,同时来自于鼻咽癌细胞的外泌体中miR-205高表达。
Objective:To investigate the role of exosome-derived miR-205 in the progression of nasopharynx carcinoma(NPC).Methods:The kit extraction method was used to extract exosomes from SUNE-1-5-8 F,SUNE-1-6-10 B and SUNE-1 NPC cell lines and NP69 normal nasopharynx epithelial cells.The morphology of exosomes was detected by electron microscope.The exosomal markers CD63 and Flotillin protein were detected by Western blot.The expression of miR-205 in exosomes was detected by real-time quantitative PCR.Results:Elliptical or rounded vesicles were separated from SUNE-1-5-8 F,SUNE-1-6-10 B and SUNE-1 NPC cell lines.The vesicles isolated from SUNE-1-5-8 F cells were uneven in size and had a complete envelope,with a diameter of 40-130 nm.The vesicles isolated from SUNE-1-6-10 B and SUNE-1 cells were uniform in size,had many electronic compact substances and a complete envelope,and with a diameter of 80-100 nm.The expression levels of CD63 and flotillin in exosomes derived from SUNE-1-5-8 F,SUNE-1-6-10 B and SUNE-1 cells were significantly higher than those in exosomes derived from NP69 cells(P<0.05).The expression level of miR-205 in exosomes derived from SUNE-1 cells was significantly higher than that in NP69 cells(P<0.05).Conclusions:Exosome is extracted and identified from NPC cells.And miR-205 is highly expressed in exosomes derived from NPC cells.
作者
汪庚明
郭术楠
张雷
周燕
营巧玲
丁建明
张亚军
江浩
项平
WANG Gengming;GUO Shunan;ZHANG Lei;ZHOU Yan;YING Qiaoling;DING Jianming;ZHANG Yajun;JIANG Hao;XIANG Ping(Department of Radiation Oncology,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Anhui Province Key Laboratory of Translational Cancer Research;Department of Cardiothoracic Surgery,The First Affiliated Hospital of Bengbu Medical College;Central Laboratory,The First Affiliated Hospital of Bengbu Medical College)
出处
《沈阳医学院学报》
2020年第1期19-24,共6页
Journal of Shenyang Medical College
基金
蚌埠医学院自然科学基金重点项目(No.BYKY1733ZD)
安徽省高校自然科学研究重点项目(No.KJ2019A0359)
蚌埠医学院转化医学重点专项项目(No.BYTM2019029)。
