【摘要】 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。更多还原

【Abstract】 Objective To develop an effective method for extracting exosomes from ascites of advanced gastric cancer patients and explore its genesis and molecular expression. Methods Ascites were collected from 4 advanced gastric cancer patients with peritoneal carcinomatosis. Exosomes were isolated by ultracentrifugation and ultrafiltration,and then examined with electron microscope. The expressions of CD63,CEA,FasL,TraiL,and HLA-G were examined with Western blotting. Results The exosomes were spherical or ovary with a diameter range of 30 to 150 nm. Exosomes from malignant ascites were positive for expressions of CD63,CEA,FasL,and Trail,but negative for HLA-G expression,while exosomes form serum of healthy volunteer were only positive for CD63 expression. Conclusion Exosomes expressing FasL and Trail can be obtained from ascites of advanced gastric cancer patients,which might play an inhibitory effect on peritoneal immunity.更多还原