SACC-83来源的外泌体调节唾液腺间质成纤维细胞表达FAP的实验研究
发布时间：2018-11-08 14:54
【摘要】：目的:研究腺样囊性癌(adenoid cystic carcinoma,ACC)细胞SACC-83来源的外泌体(exosome,EXO)对人正常唾液腺间质成纤维细胞(human normal salivary gland stromal fibroblasts,hSGSFs)表达成纤维细胞活化蛋白(fibroblast activation protein,FAP)的影响。方法:采用超滤管浓缩与EXO提取试剂盒相结合的方法从SACC-83的培养上清中提取EXO,通过透射电镜及Western Blot对所提取的EXO进行鉴定;将SACC-83来源的EXO用荧光染料PKH67标记后与hSGSFs共培养48 h,采用激光扫描共聚焦显微镜(LSCM)观察hSGSFs对于SACC-83来源的EXO的摄取情况;利用qRT-PCR和Western Blot法检测在SACC-83来源的EXO作用下,hSGSFs中FAP的表达变化。结果:SACC-83培养上清中所提取到的微囊泡直径为30~100 nm,EXO的膜蛋白标记物CD63和TSG101的表达为阳性;携带PKH67荧光标记的EXO可被hSGSFs摄取,并可在mRNA和蛋白水平上调hSGSFs中FAP的表达。结论:SACC-83来源的EXO可被hSGSFs摄取,并可显著上调hSGSFs中FAP的表达。提示ACC可能通过EXO途径促进正常唾液腺间质成纤维细胞向癌相关成纤维细胞(cancer-associated fibroblasts,CAF)的转化。
[Abstract]:Objective: to study the expression of fibroblast activating protein (fibroblast activation protein, (fibroblast activation protein,) in human salivary gland interstitial fibroblasts (human normal salivary gland stromal fibroblasts,hSGSFs) by SACC-83 derived exocrine (exosome,EXO) from adenoid cystic carcinoma (adenoid cystic carcinoma,ACC) cells. FAP). Methods: ultrafiltration tube concentration and EXO extraction kit were used to extract EXO, from the culture supernatant of SACC-83. The extracted EXO was identified by TEM and Western Blot. The EXO derived from SACC-83 was labeled with fluorescent dye PKH67 and co-cultured with hSGSFs for 48 h. The uptake of SACC-83 EXO by hSGSFs was observed by (LSCM) with laser scanning confocal microscope. The expression of FAP in hSGSFs was detected by qRT-PCR and Western Blot under the action of EXO from SACC-83. Results: the expression of CD63 and TSG101, the membrane protein markers with a diameter of 30 ~ 100 nm,EXO extracted from the supernatant of SACC-83 culture, was positive, and the EXO with PKH67 fluorescent labeling could be taken up by hSGSFs, and the FAP expression in hSGSFs could be upregulated at the mRNA and protein levels. Conclusion: EXO derived from SACC-83 can be ingested by hSGSFs and can significantly up-regulate the expression of FAP in hSGSFs. The results suggest that ACC may promote the transformation of normal salivary gland interstitial fibroblasts into cancer-related fibroblasts (cancer-associated fibroblasts,CAF) through EXO pathway.
【作者单位】： 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;
【基金】：广东省科技计划项目(编号:2014A020212397)
【分类号】：R739.8
[Abstract]:Objective: to study the expression of fibroblast activating protein (fibroblast activation protein, (fibroblast activation protein,) in human salivary gland interstitial fibroblasts (human normal salivary gland stromal fibroblasts,hSGSFs) by SACC-83 derived exocrine (exosome,EXO) from adenoid cystic carcinoma (adenoid cystic carcinoma,ACC) cells. FAP). Methods: ultrafiltration tube concentration and EXO extraction kit were used to extract EXO, from the culture supernatant of SACC-83. The extracted EXO was identified by TEM and Western Blot. The EXO derived from SACC-83 was labeled with fluorescent dye PKH67 and co-cultured with hSGSFs for 48 h. The uptake of SACC-83 EXO by hSGSFs was observed by (LSCM) with laser scanning confocal microscope. The expression of FAP in hSGSFs was detected by qRT-PCR and Western Blot under the action of EXO from SACC-83. Results: the expression of CD63 and TSG101, the membrane protein markers with a diameter of 30 ~ 100 nm,EXO extracted from the supernatant of SACC-83 culture, was positive, and the EXO with PKH67 fluorescent labeling could be taken up by hSGSFs, and the FAP expression in hSGSFs could be upregulated at the mRNA and protein levels. Conclusion: EXO derived from SACC-83 can be ingested by hSGSFs and can significantly up-regulate the expression of FAP in hSGSFs. The results suggest that ACC may promote the transformation of normal salivary gland interstitial fibroblasts into cancer-related fibroblasts (cancer-associated fibroblasts,CAF) through EXO pathway.
【作者单位】： 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;
【基金】：广东省科技计划项目(编号:2014A020212397)
【分类号】：R739.8
【相似文献】
相关期刊论文 前10条
1 饶寒敏;汤雪明;柴本甫;李慧;朱雅萍;;丹参对成纤维细胞成骨的扫描电镜观察[J];中国中西医结合杂志;2001年S1期
2 朱红梅;成纤维细胞对肿瘤发生发展的作用研究[J];医学综述;2002年06期
3 魏玉玲,邹庆,梁克玉;中药增骨Ⅰ号对体外成纤维细胞的影响[J];中医正骨;2004年01期
4 卢旭华,陈德玉,赵定麟;成纤维细胞成骨潜能的研究进展[J];骨与关节损伤杂志;2004年08期
5 魏海峰;井玲;魏雁虹;刘辉;齐玲;赵志涛;张秀云;李才;;氟对二维和三维培养的成纤维细胞胰岛素样生长因子-1表达的影响[J];中国生物制品学杂志;2010年05期
6 张诚s，
本文编号：2318815
本文链接：https://www.wllwen.com/yixuelunwen/kouq/2318815.html
最近更新
教材专著
