SACC-83-derived exosomes induce fibroblast activation protein expression in normal salivary gland stromal fibroblasts / 实用口腔医学杂志

ABSTRACT

Objective:To study the effects of exosomes (EXO) released by adenoid cystic carcinoma SACC-83 cells on the expression of fibroblast activation protein (FAP) in normal human salivary gland stromal fibroblasts (hSGSFs).Methods:ACC exosomes were isolated from SACC-83 cell culture supernatant by using Total Exosome Isolation Reagent.The whole-mount EXO were characterized and assessed by transmission electron microscope and Western Blot.The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with hSGSFs for 48h,followed by staining with Alexa Fluor 594 Phalloidin and DAPI.Mterwards,exsosomes uptake was observed under a laser scanning confocal microscope.After a 48-hour co-culture of SACC-83 exosomes with hSGSFs,the expression of FAP in SACC-83-EXO-treated hSGSFs was investigated by qRT-PCR and Western Blot.Results:The vesicles isolated from SACC-83 cell culture supernatant had the reported size range of 30-100 nm,expressed the exosomal marker CD63 and TSG101.Mter co-culture of hSGSFs with PKH67 labeled SACC-83 exosomes,exosomes were taken up by hSGSFs and FAP expression was elevated in hSGSFs.Conclusion:Exosomes derived from SACC-83 cells can be taken up by hSGSFs and can induce the expression of FAP in hSGSFs.These results suggest that exosomes derived from SACC-83 cells might induce the transformation of normal salivary gland strormal fibroblasts to cancer associated fibroblasts.