三氧化二砷抑制鼻咽癌细胞eb病毒潜伏膜蛋白1及其抗癌效应的研究-study on arsenic trioxide inhibiting latent membrane protein 1 of nasopharyngeal carcinoma cell eb virus and its anti-cancer effect.docxVIP

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三氧化二砷抑制鼻咽癌细胞eb病毒潜伏膜蛋白1及其抗癌效应的研究-study on arsenic trioxide inhibiting latent membrane protein 1 of nasopharyngeal carcinoma cell eb virus and its anti-cancer effect.docx

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三氧化二砷抑制鼻咽癌细胞eb病毒潜伏膜蛋白1及其抗癌效应的研究-study on arsenic trioxide inhibiting latent membrane protein 1 of nasopharyngeal carcinoma cell eb virus and its anti-cancer effect

PAGE PAGE 10 ABSTRACT Aims and background: It was demonstrated that nasopharyngeal carcinoma (NPC) is closely related to Epstein-Barr virus (EBV) infection and the virus encoded latent membrane protein-1 (LMP1) takes an important role in the pathogenesis of NPC, blocking differentiation and against apoptosis as well as promoting metastasis. In our preclinical study, As2O3 has been implicated as a promising anticancer agent in inducing apoptosis and differentiation of NPC xenografts in Scid mouse. However, whether the mechanism is involved in the repression of LMP1 remained unknown. Furthermore, the role of changes of LMP1 expression influenced by As2O3 in NPC cells in indicating apoptosis and invasion potential also need to be investigated. Therefore, the thesis consisting of two parts is designed to answer the above questions. In part one, the purpose is to study whether As2O3 can inhibit LMP1 expression in HNE1-LMP1 cells. Then, the cell telomere length alteration and apoptosis are detected in order to explore the role of LMP1 inhibition in the apoptosis effect mediated by As2O3. The aim of the part two is to observe the metastasis potential of residual cells after treated by As2O3 and its possible mechanisms. The relationship between LMP1 status and MMP9 was also analyzed in attempt to explore the role of LMP1 in this procedure. Methods: Stable expression of LMP1-positive human NPC cell line HNE1-LMP1 and its parental LMP1-negative cell line HNE1 were used as in vitro models. In the part one, HNE1-LMP1 cells (1 × 105/mL) were cultured in the media containing varied concentration of As2O3 (0、1、2、3、4、 5μmol/L )for 48 hours, respectively. Western-blot and the laser scanning confocal microscopy were performed to observe the changes of latent membrane protein 1 (LMP1) expression. The level of mRNA was also detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay.Then, HNE1-LMP1’s parental cell HNE1 is used as control to compare t