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目的从胃癌患者腹水中分离exosomes并鉴定其来源及分子表型。方法采用多步超速离心加超滤方法从胃癌患者腹水中分离exosomes,采用透射电镜观察其形态,采用Wenstern blot检测其CD63、FasL、TraiL、HLA-G等分子的表达。结果从胃癌患者腹水中分离的exosomes形态为圆形或椭圆形小囊泡,直径约30～100nm;经Western blot证实,exosomes表达CD63、CEA、FasL、TraiL等分子,不表达HLA-G。正常血清来源的exosomes只表达CD63。结论从胃癌患者腹水中分离的exosomes来源于胃癌细胞,表达FasL及TraiL,可能对腹膜腔的免疫细胞起到抑制作用。
Objective To isolate exosomes from ascites of patients with gastric cancer and identify their origin and molecular phenotype. Methods Exosomes were isolated from ascites of gastric cancer patients by multi-step ultracentrifugation and ultrafiltration. Morphological changes were observed by transmission electron microscopy. The expression of CD63, FasL, TraiL, HLA-G and other molecules were detected by Wenstern blot. Results The morphology of exosomes isolated from ascites of gastric cancer patients was round or oval vesicles with a diameter of 30 ~ 100 nm. Western blot confirmed that exosomes expressed CD63, CEA, FasL and TraiL molecules but not HLA-G. Normal serum derived exosomes only express CD63. Conclusion The exosomes isolated from ascites in patients with gastric cancer originate from gastric cancer cells and express FasL and TraiL, which may inhibit the peritoneal immune cells.