Analyzing the HPV E6/E7 Oncoproteins in Human Cervical Cancer Specimens

• Main Authors: Hsueh-Ting Huang, 黃雪婷
• Other Authors: 林如華
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/51354449996983539947

碩士 === 中國醫藥大學 === 生物科技學系碩士班 === 97 === In 2007, the Department of Health information shows that cervical cancer is the sixth cause of female cancer death in Taiwan. It’s still the second most common malignancy and the second most common cause of cancer-related death in women worldwide from WHO information. Human papillomavirus has been closely linked with human cervical cancer. The objectives of this study are to 1). detect HPV E6 and E7 proteins expressed in cervical cancer cell lines using polyclonal and monoclonal antibody we generated specific to the HPV oncoproteins. 2). develop immunoassays for detection of HPV E6, E7 oncoproteins using pap smear samples (liquid based). 3). compare the results of HPV DNA test, immunoassays, and clinical diagnosis (histology or pap smear results). We collected cervical specimens from 204 women with cervix cell and 12 women with cervical cancer tissue. Our preliminary results show that HPV E6 and E7 oncoproteins are detected from cervical cancer cell lines such as HeLa (HPV18), CaSki (HPV16), but not in C33A (HPV negative) by western blot using the specific HPV antibody. ELISA data indicate direct detection of HPV E7 oncoprotein from various stages of cervical scrape samples. Compared ELISA results to PCR, high correlation was found in these two methods. Immunocytochemistry assay also demonstrates the overexpression of E6 and E7 oncoproteins present in the nuclear of abnormal cells from various stage of liquid based cervical scrape samples. Therefore, various formats, high throughput, user friendly immunoassays can be further developed.