人正常唾液腺上皮细胞摄取腺样囊性癌细胞来源的外泌体的实验研究

中国口腔颌面外科杂志 ›› 2016, Vol. 14 ›› Issue (2): 106-110.

人正常唾液腺上皮细胞摄取腺样囊性癌细胞来源的外泌体的实验研究

宋梦阳, 杨小军, 侯晋, 殷学民

南方医科大学南方医院口腔颌面外科,南方医科大学口腔医学院,广东广州 510515

收稿日期:2015-09-23 出版日期:2016-03-20 发布日期:2016-04-06
通讯作者: 殷学民,E-mail:yinxm@fimmu.com
作者简介:宋梦阳（1991-）,女,硕士,E-mail:songmengyang0913@126.com
基金资助:
广东省科技计划项目（2014A020212397）

Exosomes from adenoid cystic carcinoma cells can be uptaken by normal human salivary gland epithelial cells

SONG Meng-yang, YANG Xiao-jun, HOU Jin, YIN Xue-min

Department of Oral and Maxillofacial Surgery, Nanfang Hospital;College of Stomatology, Southern Medical University. Guangzhou 510515, Guangdong Province, China

Received:2015-09-23 Online:2016-03-20 Published:2016-04-06

摘要/Abstract

摘要： 目的:研究腺样囊性癌ACC-2细胞来源的外泌体能否靶向正常唾液腺上皮细胞（SGECs）,为进一步研究ACC来源的外泌体在ACC侵袭和转移中的调节作用奠定基础。方法:通过组织块法原代培养人SGECs,应用免疫细胞化学染色对其来源进行鉴定;将ACC-2细胞来源的外泌体用荧光染料PKH67标记后与SGECs共培养,用扫描共聚焦显微镜(LSCM)观察SGECs对ACC-2来源的外泌体的摄取情况。结果:原代培养的SGECs高表达角蛋白CK19,不表达α-MSA,说明其来源可能为腺泡上皮或导管上皮细胞,而非间质和肌上皮细胞。在LSCM下观察到携带PKH67标记的外泌体可以进入SGECs内,主要分布于核周的胞质中。结论:SGECs可摄取ACC-2细胞来源的外泌体,提示可能为ACC来源外泌体的靶细胞之一。

关键词: 腺样囊性癌, 外泌体, 唾液腺上皮细胞

Abstract: PURPOSE: To detect whether human salivary gland epithelial cells (SGECs) can uptake exosomes derived from adenoid cystic carcinoma cell ACC-2. METHODS: Human normal SGECs were primarily cultured by explant out growth technique and identified by immunocytochemistry. The exosomes derived from ACC-2 were labeled with green fluorescent dye PKH67 and co-cultured with SGECs for 48h, which were then stained with Alexa Fluor 594 Phalloidin and DAPI. Afterwards, exsosomes absorption was observed under a laser scanning confocal microscope (LSCM). RESULTS: Primarily cultured normal human SGECs exhibited strong cytokeratin expression but absence of staining for α-MSA, indicating the lack of myoepithelial cells in the culture system; after 48 h of incubation, multiple green vesicles could be seen docked on SGECs and most diffuse PKH67 signals were concentrated in the cytoplasm. Moreover, the uptake of exosomes was highly efficient as all SGECs were positive for stained exosomes. CONCLUSIONS: Normal human SGECs could uptake ACC-2 derived-exosomes, indicating that SGECs might be a kind of recipient cells of ACC-2 derived- exosomes.

Key words: Adenoid cystic carcinoma, Exosomes, Salivary gland epithelial cells

中图分类号:

R739.8

宋梦阳, 杨小军, 侯晋, 殷学民. 人正常唾液腺上皮细胞摄取腺样囊性癌细胞来源的外泌体的实验研究[J]. 中国口腔颌面外科杂志, 2016, 14(2): 106-110.

SONG Meng-yang, YANG Xiao-jun, HOU Jin, YIN Xue-min. Exosomes from adenoid cystic carcinoma cells can be uptaken by normal human salivary gland epithelial cells[J]. China J Oral Maxillofac Surg, 2016, 14(2): 106-110.

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