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基于双向电泳荧光差异凝胶电泳和基质辅助激光解吸电离飞行时间质谱技术的卵巢上皮癌血清生物标志物分析

[Analysis of serum biomarkers of ovarian epithelial cancers based on 2-DE DIGE and MALDI TOF/TOF].

作者信息

Zhao Qun, Duan Wei, Wu Yu-mei, Qian Xiao-hong, Deng Xiao-hong

机构信息

Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2008 Oct;30(10):754-8.

Abstract

OBJECTIVE

To find new serum tumor markers for ovarian epithelial cancers by 2-DE DIGE and MALDI-TOF/TOF proteomic methods, in order to improve the diagnostic sensitivity and specificity.

METHODS

Serum samples from 103 cases of ovarian epithelial cancers, 60 cases of healthy women, 63 cases of benign ovarian tumors and 63 cases 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 sera (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.

摘要

目的

通过二维差异凝胶电泳(2-DE DIGE)和基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF)蛋白质组学方法寻找卵巢上皮癌新的血清肿瘤标志物,以提高诊断的敏感性和特异性。

方法

收集103例卵巢上皮癌患者、60例健康女性、63例卵巢良性肿瘤患者及63例盆腔良性疾病患者的血清样本。将20例卵巢上皮癌患者(A组)、20例卵巢良性肿瘤患者(B组)、20例盆腔良性疾病患者(C组)及20例健康对照者(D组)的血清按年龄匹配后分别混合。去除高丰度血清白蛋白和免疫球蛋白G后,采用2-DE DIGE对样本进行检测。该检测重复3次。用DeCyder软件分析显示有显著差异的蛋白质点,并用MALDI-TOF/TOF进行鉴定。采用蛋白质免疫印迹法(Western blot)和酶联免疫吸附测定法(ELISA)对候选血清标志物进行验证。

结果

1)各组间有41种蛋白质存在显著差异。MALDI-TOF/TOF成功鉴定出28种蛋白质。触珠蛋白(Hp)是上调最显著的蛋白质,转铁蛋白(Tf)是下调最显著的蛋白质。2)蛋白质免疫印迹法和酶联免疫吸附测定法证明,卵巢上皮癌与正常对照之间(P = 0.000)、卵巢上皮癌与卵巢良性肿瘤之间(P = 0.000)、卵巢上皮癌与盆腔良性疾病血清之间(P = 0.000),Hp和Tf均存在显著差异。3)CA125 + Hp + Tf联合检测卵巢癌的敏感性和特异性高于单独检测CA125、Hp或Tf。

结论

Hp和Tf在卵巢上皮癌患者血清中表达不同。它们可作为卵巢上皮癌的血清生物标志物。CA125 + Hp + Tf联合检测可能提高卵巢上皮癌诊断的敏感性和特异性。

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