针对 HPV16 E7 和 GP96 片段的抗体检测作为宫颈癌患者的生物标志物。

Antibody detection against HPV16 E7 & GP96 fragments as biomarkers in cervical cancer patients.

机构信息

Molecular Immunology & Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran.

出版信息

Indian J Med Res. 2009 Nov;130(5):533-41.

PMID:20090101

Abstract

BACKGROUND & OBJECTIVE: Cervical cancer is the second most frequent cancer among females worldwide, especially human papilloma viruses (HPV) types 16 and 18. In viral systems the identification of serological markers would facilitate the diagnosis of HPV infections and virus-related disease. The aim of the present investigation was to determine and search for serologic markers in cervical cancer patients associated with HPV.

METHODS

A total of 58 Iranian women with invasive cervical carcinoma including adenocarcinoma and squamous cell carcinoma (SCC) were included. Serum antibody response to HPV infections in patients was detected by Western blot and ELISA techniques based on recombinant HPV16E7 and the N-terminal and C-terminal fragments of gp96 (NT-gp96 and CT-gp96) proteins. These recombinant proteins were expressed in Escherichia coli as a His-tag protein and purified using affinity chromatography.

RESULTS

The ELISA results indicated that patients with high antibody response to HPV16E7 had significant seroreactivity to CT-gp96 fragment. In Western blot analysis, a strong association between anti-E7, anti-NT-gp96 and anti-CT-gp96 reactivity and cervical cancer was obtained using purified recombinant proteins. In adenocarcinoma cases, no significant difference was observed in seroreactivities between normal and patients.

INTERPRETATION & CONCLUSION: The evaluation of cervical cancer patients' seroreactivities against three recombinant proteins (rE7, rNT-gp96 and rCT-gp96) showed significantly higher levels of these markers in SCC only, but not in adenocarcinoma and control groups. Also, the usage of both techniques (ELISA and Western blotting) can provide more reliable tools for diagnosis of cervical cancer.

摘要

背景与目的

宫颈癌是全世界女性中第二常见的癌症，尤其是人乳头瘤病毒（HPV）16 型和 18 型。在病毒系统中，鉴定血清标志物将有助于 HPV 感染和与病毒相关疾病的诊断。本研究旨在确定和寻找与 HPV 相关的宫颈癌患者的血清学标志物。

方法

共纳入 58 例伊朗浸润性宫颈癌患者，包括腺癌和鳞状细胞癌（SCC）。通过 Western blot 和 ELISA 技术检测患者对 HPV 感染的血清抗体反应，该技术基于 HPV16E7 重组蛋白和 gp96 的 N 端和 C 端片段（NT-gp96 和 CT-gp96）。这些重组蛋白在大肠杆菌中作为 His 标签蛋白表达，并通过亲和层析进行纯化。

结果

ELISA 结果表明，对 HPV16E7 具有高抗体反应的患者对 CT-gp96 片段具有显著的血清反应性。在 Western blot 分析中，使用纯化的重组蛋白获得了抗-E7、抗-NT-gp96 和抗-CT-gp96 反应与宫颈癌之间的强烈关联。在腺癌病例中，正常组和患者组之间的血清反应性没有显著差异。