摘要目的 通过差异蛋白质组学的方法寻找卵巢上皮癌血清标志物,以提高卵巢癌诊断的灵敏度和特异度.方法 收集103例卵巢上皮癌、60例正常对照、63例卵巢良性肿瘤和63例盆腔良性病变患者的血清.按照患者年龄匹配,选取卵巢上皮癌、盆腔良性病变、卵巢良性肿瘤和正常对照者血清各20例,等容积混合,去除高丰度白蛋白和IgC后,进行荧光双向电泳(2-DE DIGE),实验重复3次.以OyCyder软件分析得出有显著差异的蛋白质点,以基质辅助激光解吸离子飞行时间串联质谱(MALDI TOF/TOF)鉴定差异蛋白质,分别采用Western blot和酶联免疫吸附实验(ELISA)验证获选的血清标志物.结果 2-DE DIGE得出有显著差异的蛋白质点41个,MALDI TOF/TOF成功鉴定出其中的28个蛋白质.上调最显著的蛋白质为结合珠蛋白(Hp),下调最显著的蛋白质为转铁蛋白(Tf).Western blot和ELISA验证结果显示,Hp和Tf在卵巢上皮癌与正常对照、卵巢上皮癌与卵巢良性肿瘤、卵巢上皮癌与盆腔良性病变之间的差异有统计学意义(均P=0.000).CAl25+Hp+Tf联合检测诊断卵巢上皮癌的灵敏度(81.6%)和特异度(76.9%)高于CA125(78.6%和70.4%)、Hp(69.9%和70.4%)和Tf(66.0%和70.4%)单独检测.结论 Hp和Tf是卵巢上皮癌血清中差异表达的蛋白质,可作为卵巢上皮癌的血清标志物,CAl25+Hp+Tf联合检测有助于提高卵巢上皮癌诊断的灵敏度和特异度.

abstractsObjective To find new serum tumor markers for ovarian epithelial cancers by 2-DE DIGE and MALDI-TOF/TOF proteomie methods, in order to improve the diagnostic sensitivity and specificity. Methods Serum samples from 103 eases of ovarian epithelial cancers, 60 eases of healthy women, 63 eases of benign ovarian tumors and 63 eases of benign pelvic diseases were collected. Sera of 20 cases of ovarian epithelial cancers (A), 20 cases of ovarian benign tumors (B), 20 cases of pelvic benign diseases (C) and 20 cases of health control (D) were matched by age and pooled, respectively. After depletion of high abundance serum albumin and IgG, the samples were assayed by 2-DE DIGE. The test was repeated three times. Analysis with DeCyder software revealed significant differential protein spots which were identified by MAIDI-TOF/TOF. Western blot and ELISA were used to validate the candidate serum markers. Results 1) There were 41 proteins having significant differences between the groups. MAIDI-TOF/TOF successfully identified 28 proteins. Haptoglobin (Hp) was the most significantly up-regulated protein, and transferrin (Tf) was the most significantly down-regulated protein. 2) Western blot and ELISA proved that there were significant differences in Hp and Tf between ovarian epithelial cancers and normal controls (P=0.000), between ovarian epithelial cancers and ovarian benign tumors (P=0.000), between ovarian epithelial cancers and benign pelvic disease sere (P=0.000).3) CA125 + Hp + Tf combined detection of ovarian cancer had higher sensitivity and specificity than CA125, Hp or Tf detection alone. Conclusion Hp and Tf are differently expressed in the sera of patients with ovarian epitheliual cancers. They can be used as serum biomarkers for ovarian epithelial cancers. CA125 + Hp + Tf combined detection may improve the sensitivity and specificity of diagnosis of ovarian epithelial cancers.