| 不同转移习性人鼻咽癌细胞株中外泌体的提取及鉴定 |
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| 引用本文： | 郭术楠,汪庚明,张雷,营巧玲,丁建明,周燕,周育夫,江浩. 不同转移习性人鼻咽癌细胞株中外泌体的提取及鉴定[J]. 中华全科医学, 2020, 18(8): 1276. DOI: 10.16766/j.cnki.issn.1674-4152.001484 |
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| 作者姓名： | 郭术楠 汪庚明 张雷 营巧玲 丁建明 周燕 周育夫 江浩 |
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| 作者单位： | 1. 蚌埠医学院第一附属医院肿瘤放疗科, 安徽 蚌埠 233004; |
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| 基金项目： | 安徽省高校自然科学研究重点项目(KJ2019A0359)蚌埠医学院自然科学基金重点项目(BYKY1733ZD)蚌埠医学院转化医学重点专项项目(BYTM2019029) |
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| 摘 要： | 目的 探索不同转移习性人鼻咽癌细胞株SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1和正常鼻咽上皮细胞NP69细胞外泌体的表达情况,为进一步研究鼻咽癌的诊断和治疗新方法提供一定的理论基础。 方法 在2018年5月—2019年6月期间,用Exosome提取试剂盒从SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1和NP69培养上清液分离出外泌体,使用透射电镜进行观察验证外泌体形态,BCA试剂盒测定外泌体蛋白的浓度,蛋白质印迹法(Western blotting)法进行外泌体特异性抗原分子CD63及筏蛋白-1的检测。 结果 3种鼻咽癌细胞系和正常鼻咽上皮细胞上清液中均分离出泡状物,在透射电镜下呈椭圆形或者圆形的泡状物。SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1和NP69细胞系外泌体蛋白浓度分别为0.778、0.635、0.788、0.576μg/μL;且所有细胞系来源的外泌体经检测均表达特异性抗原分子CD63及筏蛋白-1。 结论 3种鼻咽癌细胞株SUNE-1-5-8F、SUNE-1-6-10B、SUNE-1和正常鼻咽上皮细胞NP69细胞均产生外泌体。
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| 关 键 词： | 鼻咽肿瘤 外泌体 鉴定 |
| 收稿时间： | 2019-10-14 |
Extraction and identification of exosomes from human nasopharyngeal carcinoma cell lines with different metastatic habits |
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| Affiliation: | Department of Radiation Oncology,the First Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui 233004,China |
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| Abstract: | Objective To explore the expression of exosomes in human nasopharyngeal carcinoma(NPC) cell lines SUNE-1-5-8 F, SUNE-1-6-10 B, SUNE-1 and normal nasopharyngeal epithelial cells NP69, and to provide a theoretical basis for further study of new methods of diagnosis and treatment of NPC. Methods Exosomes were isolated from SUNE-1-5-8 F, SUNE-1-6-10 B, SUNE-1 and NP69 culture supernatants using exosome extraction kits, and verified by transmission electron microscopy. The concentration of exosome protein was determined by BCA kit. Eexosome specific antigen molecule CD63 and Flotillin-1 were detected by western bolt. Results Vesicles were isolated from supernatant of three kinds of nasopharyngeal carcinoma cell lines and normal nasopharyngeal epithelial cells, showed a circular vesicle under a transmission electron microscope. The exosomal protein concentrations of SUNE-1-5-8 F, SUNE-1-6-10 B, SUNE-1 and NP69 were 0.778, 0.635, 0.788, and 0.576 μg/μL, respectively. CD63 and Flotillin-1 were expressed in all cell lineage-derived exosomes. Conclusion Three kinds of nasopharyngeal carcinoma cell lines SUNE-1-5-8 F, SUNE-1-6-10 B, SUNE-1 and normal nasopharyngeal epithelial cell line NP69 all produce exosomes. |
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