Detections of the HPV-18 E6 Antigen and Anti-E6 Antibody in Human Cervical Cancer

• Main Authors: Chan-Yen Kuo, 郭展延
• Other Authors: Guang-Yuh Hwang
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/36511303709215799876

碩士 === 東海大學 === 生物學系 === 92 === Cervical cancer is the most common cancer in women worldwide. There is a strong epidemiological correlation between human papillomavirus (HPV) infections and cervical cancer. The information on the serological studies on the patient with cervical cancer caused by HPV was very limited. In this study, the serological aspects of the HPV-18 E6 antigen, the viral nonstructural protein, were approached. First of all, the 25 patients’ tissues with cervical cancer were obtained and the E6 antigen extracted from cervical cancer tissues and their counter parts were specifically detected by anti-E6 monoclonal antibody. Furthermore, the E6 antigen expressed in patients’ cervical tissues with cervical cancer and normal counter parts using anti-E6 monoclonal antibody was detected by immunological staining. These clinically-confirmed patients with the molecular serotyping were used for serological studies. To measure the titers of sera from patients with cervical cancer, the HPV-18 E6 was expressed in E. coli DH5α by recombinant DNA technology using glutathione S-transferase (GST) fusion system, and purified to homogeneity by glutathione-sepharose 4B beads. Immunological characterizations of the fusion protein were studied by western immunodetection. The specific bindings between the anti-HPV E6 monoclonal antibody and the recombinant HPV E6 was shown. These data show the precision of the recombinant HPV E6 fusion protein. The recombinant nonstructural E6 fusion protein and the purified E6 protein were used to screen the titers of anti- E6 antibody in sera. Titers of anti-E6 antibody in sera from twenty-five patients with cervical cancer or twenty-two normal individuals were evaluated by quantitative ELISA. Results indicate that the sera of patients with cervical cancer show significant binding to the fusion protein GST-E6 and the purified E6. Furthermore, each of the twenty-five patients’ sera was also reacted to the same amount of GST-E6 proteins. The titers of anti-E6 antibody show stronger binding than the anti-L1 antibody among all 25 tested sera. These results indicate that the titer of antibody against the viral nonstructural protein E6 was elevated in sera from patients with cervical cancer.