2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38155Epstein-Barr virus (EBV) is a human herpesvirus, which is persistent in B lymphoid cells, triggers transformation and tumorigenesis of latently infected B cells in vitro via LMP-induced cellular survival protein, Bcl-2. In other words, Bcl-2 protein in EBV-infected B cells plays an important role in prolonging life span of cells. Recently, an early lytic gene, BALF1, which is structurally homologous to Bcl-2 protein, is characterized in EBV infected cells. According to the preliminary results, we have known that BALF1 inhibited EBNA1 transcript and was involved in repressing the cell apoptosis through the ceramide-mediated signal pathway. However, the function of BALF1on EBV life cycle and cell apoptosis still needs to be clarified. In this proposal, we initially intend to establish a BALF1 stable cell line, which stably expresses BALF1 protein and use this stable cell line to observe the effect of BALF1 on the promoters of latent genes (LMP and EBNA1) and immediate early genes, Z promoter. In the meantime, we also intend to test the impact of BALF1 on EBV production and cell transformation. In addition, we will attempt to perform DNA repair assay and detect the expression of tumor suppressor gene to demonstrate involvement of BALF1 in cell tumorigenesis. On the other hand, we will monitor the activity or amount of JNK at post transfection of BALF1 to explore the relationship between BALF1 and ceramede-mediated apoptosis. Furthermore we use various apoptosis-inducing agents to clarify the effect of BALF1 on the other pathway of cell apoptosis. Finally we will employ human cellular cDNA microarray to find out which cellular genes that involved in cell apoptosis, tumorigenesis and growth are affected by BALF1. The results of microarray assay will provide us more information that how BALF1 regulates cell tumorigenesis in EBV infected cells. From this project, we will obtain information what function of BALF1 is in infected cells and how to regulate EBV life cycle and host cells. Understanding the function of BALF1 in infected cells will facilitate the development of drugs to inhibit its intrinsic activities and non-immunogenic vectors for use in human gene therapy.EB病毒是人類皰療病毒(herpesvirus)的一員，它能夠持續存活於B淋巴細胞，並透 過其潛伏性膜蛋白1 (LMP1)活化細胞中的Bcl-2蛋白質。導致B細胞轉形(transformation) 及癌化(tumorigenesis)。換言之，Bcl-2蛋白在其中扮演非常重要的角色。近來在EB病 毒研究中，發現有一種會出現在溶裂期早期的蛋白質，其結構與Bcl-2相似，我們稱它 爲BALF1。根據我們初步研究結果，已知BALF1具有抑制EB病毒EBNA1的transcript 及ceramide所引起的細胞调亡(apoptosis)的能力°然而至今對於它在細胞中的主要功能 仍存有許多不清之處。爲了釐清此點，在本計劃中首先建立穩定表現BALF1蛋白的細 胞株(cell line)，接著利用此細胞株觀察BALF1對EB病毒潛伏及溶裂基因啓動子的影 響。同時，測試其對EB病毒顆粒的產生及細胞的轉形。此外，我們嘗試研究BALF1 對細胞基因體穩定所扮演的角色，並深入了解細胞癌化。在另一方面，我們偵測細胞 中JNK蛋白的表現量及活性，以便探討BALF1與ceramide引起的細胞凋亡的相關性。 也利用流式細胞儀去觀察BALF1與其他凋亡途徑的關係。最後，我們利用人類的cDNA 微矩陣(microarray)，去尋找受BALF1調控的細胞基因，進而觀察其如何改變細胞的狀 態。從本次研究計劃，我們可以從中獲得BALF1基因對其宿主及病毒本身調控機制的 資訊。而從資訊中又可找出宿主與病毒間互動的關係。藉由深入了解此關係，可發展 出抑制EB病毒的藥物及因病毒所產生疾病的基因治療方法。Project