B型肝炎病毒HBx蛋白質在人類肝腫瘤組織細胞中的存在及分佈

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题名:	B型肝炎病毒HBx蛋白質在人類肝腫瘤組織細胞中的存在及分佈
其它题名:	Identification and distribution of the hepatitis B virus X protein in human hepatocellular carcinoma cells
作者:	許靜婷 Hsu, Ching-Ting
贡献者:	黃光裕 Hwang, Guang-Yuh 東海大學生命科學系
关键词:	HBx蛋白質 HBx;hepatocellular carcinoma
日期:	2002
上传时间:	2011-04-27T05:46:51Z (UTC)
摘要:	在台灣，肝癌為十大死因之一。B型肝炎病毒所轉錄的HBx蛋白質是造成肝癌的主要原因。X基因是最小的一段B型肝炎病毒基因組的開放讀碼區，能夠合成HBx蛋白質。HBx蛋白質是一個多功能蛋白質，且與肝癌的發生有關。但是其功能上的機制卻了解得很少，雖然有幾個細胞核和細胞質間交互作用的蛋白質已於活體外被證實了，但對於HBx蛋白質於被感染的肝細胞上的分佈卻沒有一致且明確的答案。所以本研究將以B型肝炎病毒HBx蛋白質在人類肝腫瘤組織中的定量及分佈為目標。使用抗HBx蛋白質的單株抗體，以西方點墨轉漬法（western blotting），來免疫偵測萃取自20個HCC病人的肝腫瘤組織。其結果顯示有85%的肝腫瘤組織中含有專一的HBx蛋白質，且在電泳膠上會和純化的重組HBx蛋白質同步移動。為了更進一步了解HBx蛋白質的角色，故以免疫組織化學染色法來檢測HBx蛋白質在肝癌組織細胞中的分佈。在8個新鮮的肝組織樣品，包括了來自於4個肝癌病人身上正常和腫瘤的部分。HBx蛋白質在正常和肝腫瘤組織，以單株和多株的HBx蛋白質抗體均可以被偵測得到。且HBx蛋白質幾乎全部分佈於細胞質中。HBx蛋白質分佈的分析，可提供將來HBx功能研究的一個基礎。 Hepatocellular carcinoma (HCC) is one of the ten major leading death diseases in Taiwan. The causal agent of HCC is hepatitis B virus (HBV) which encodes a protein (HBx). X gene is the smallest open reading frame (ORF) among hepatitis B virus (HBV) genome and it encodes the HBV X protein (HBx). The HBx is a multifunctional protein that is implicated in the pathogenesis of hepatocellular carcinoma. Very little is known about its functional mechanisms although interactions with several nuclear and cytoplasmic proteins have been demonstrated in vitro, there is no clear consensus as to the location of HBx in infected hepatocytes. So quantification and cellular distribution of the hepatitis B virus X protein in human hepatocellular carcinoma were studied. Western immunodetection of HBx antigen in liver extracts of twenty HCC patients was performed using the anti-HBx monoclonal antibody. The results show that 85% liver tissues of HCC contain a specific and comigrated HBx antigen. To further elucidate the role of HBx, the distribution of HBx was also examined by immunohistochemical staining. Eight fresh liver tissue samples, normal and tumor parts of four HCC patients were analysed. HBx was detected in both normal and tumor tissues using anti-HBx monoclonal and polyclonal antibodies. HBx was mainly located in cytoplasm. This distribution analysis of HBx will provide the basis for the study of the functional identification of the HBx.
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