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    題名: 子宮頸癌之人類乳突瘤病毒E6/E7蛋白的表現分析;Analyzing the HPV E6/E7 Oncoproteins in Human Cervical Cancer Specimens
    作者: 黃雪婷;Hsueh-Ting Huang
    貢獻者: 中國醫藥大學:生物科技學系碩士班
    關鍵詞: 子宮頸癌;人類乳突瘤病毒;E6/E7致癌蛋白;cervical cancer;human papillomavirus;E6/E7 oncoproteins
    日期: 2009-06-23
    上傳時間: 2009-08-11 10:58:35 (UTC+8)
    摘要: 根據2007年行政院衛生署統計子宮頸癌是目前台灣地區女性癌症死因的第六位,死亡率約為每十萬名人口7.0例,在世界衛生組織的統計下更是佔全球女性常見癌症的第二位。研究發現高危險型的人類乳突瘤病毒E6/E7基因,藉由與腫瘤抑制基因的結合,促使細胞不正常增生,導致細胞癌化,為子宮頸細胞癌化的主要原因。?d了分析人類乳突瘤病毒E7/E6致癌蛋白的表現於子宮頸癌發展過程所扮演的角色,本論文嘗試建立利用抗體分析人類子宮頸細胞檢體中,人類乳突瘤病毒E6/E7致癌蛋白表現的方法,由不同階段之臨床病人檢體的分析中,了解人類乳突瘤病毒E6/E7蛋白的表現,與子宮頸癌發展的關聯性,以提供未來發展早期子宮頸癌篩檢的方法。本研究以204例正常之子宮頸刮除細胞、12例子宮頸癌患者癌化組織及子宮頸癌細胞株,於西方墨點法實驗結果顯示,所使用之人類乳突瘤病毒E6/E7抗體,可專一地偵測到子宮頸癌細胞株中的目標蛋白。在酵素連結免疫吸附分析法中,直接式酵素連結免疫吸附分析法與聚合?○s鎖反應(PCR)間結果有較高的關聯性;於細胞免疫染色結果中亦顯示,人類乳突瘤病毒 E6/E7蛋白確實存在於不正常細胞,且根據臨床診斷的分期,其表現程度有所不同。本研究結果初步證明,利用直接式酵素連結免疫吸附法與細胞免疫染色,可用於分析人類子宮頸細胞檢體中,人類乳突瘤病毒E6/E7致癌蛋白的表現。

    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.
    顯示於類別:[生物科技學系暨碩士班] 博碩士論文

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