卵巢上皮性癌患者腹水中外来体的分离和鉴定及临床意义

目的 分离和鉴定卵巢上皮性癌(卵巢癌)患者腹水中的外来体(exosome),并探讨其临床意义.方法 收集2006年3月-2007年3月间收治的41例卵巢癌患者的腹水,通过差速离心联合密度屏障离心法分离腹水中的外来体,分别采用透射电镜和蛋白印迹法对外来体进行形态学和分子标志物鉴定,并回顾性分析卵巢癌患者腹水中出现外来体的临床意义.结果 卵巢癌患者腹水中可成功分离并获得外来体,卵巢癌患者腹水中外来体的阳性率为85%(35/41);其形态具有典型的外来体的形态学特征,且表达外来体的相对特异性分子标志物,其中组织相容性复合物I(MHC-I)阳性率为100%(35/35)、热休克蛋白70(Hsp70)阳性率为91%(32/35)、CD81.阳性率为86%(30/35).不同年龄、病理类型、病理分级、手术病理分期、手术的彻底性、术后血清CA125水平下降满意度、化疗反应的卵巢癌患者腹水中外来体的阳性率比较,差异均无统计学意义(P>0.05).卵巢癌患者行肿瘤细胞减火术后,腹水中外来体阳性患者血清CA125水平平均下降幅度为(66±27)%,明显高于外来体阴性患者的(37±86)%及卵巢痛患者整体的(61±44)%(P<0.01);且这种差异与手术是否彻底无关(P<0.01).结论 卵巢癌患者腹水中能够分离到典型的外来体.外来体的存在与卵巢癌患者肿瘤细胞减灭术后CA125水平下降幅度有关,但与其预后的关系有待进一步研究.

Isolation,identification and clinical significance of ascites-derived exosomes from patients with ovarian epithelial cancer

Objective To isolate and identify the aseites-derived exosomes from patients with ovarian epithelial carcinoma,and explore the relationship between exosomes and the prognosis of ovarian epithelial carcinoma.Methods Ascites-derived egosomes were isolated by ultraeentrifugation on sucrose and D2O gradients from 41 ovarian epithelial carcinoma patients,and identified by transmission electron microscope and western blot analyses.Ascites-derived exosomes were evaluated for effect on prognosis of ovarian epithelial carcinoma.Resulls Exosomes were isolated and purified from the ascites in 85% (35/41)of ovarian epithelial carcinoma patients;major histocompability complex-I(MHC-I)could be detected in 100%(35/35)of the aseites-derived exosomes samples,heat shock protein-70(Hsp70)in 91%(32/35),and CD81 in 86%(30/35).The patients with positive ascites-derived exosomes had no significant difference in age,pathological type and the degree of differentiation of tumor,surgical-pathological staging,the optimal operation and the responsibility to chemotherapy(P>0.05).The patients with positive ascites-derived exosomes,the reduction of CA125 level after cytoreduetive surgery was(66±27)%,which was more than that of the patients without ascites-derived exosomes(37±86)%and all the whole patients not considering the condition of exosomes(61±44)%(P<0.01),and there were also no difference whether or no optimal operation ration(P<0.01).Conclusions There are ascites-derived exosomes in ascites from most patients with ovarian epithelial carcinoma.While the relationship between exosomes and prognosis of ovarian epithelial carcinoma needs further research.