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研究生:王登五
研究生(外文):Wang, Deng-Wu
論文名稱:腸病毒71型感染誘發巨噬細胞移動抑制因子於人類肺腺癌細胞與橫紋肌肉瘤細胞之表現
論文名稱(外文):Enterovirus 71 Infection Induces Expression of Macrophage Migratory Inhibitory Factor in Human Lung Adenocarcinoma Cells and Rhabdomyosarcoma Cells
指導教授:傅在峰
指導教授(外文):Fu, Tsai-Feng
口試委員:陳連城吳立真
口試委員(外文):Chen, Lien-ChengWu, Li-Chen
口試日期:2012/07/23
學位類別:碩士
校院名稱:國立暨南國際大學
系所名稱:應用化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:英文
論文頁數:62
中文關鍵詞:腸病毒71型巨噬細胞移動抑制因子橫紋肌肉瘤細胞人類肺腺癌細胞缺氧誘發因子高遷移率族蛋白
外文關鍵詞:Enterovirus 71macrophage migration inhibitory factorrhabdomyosarcoma cellhuman lung adenocarcinoma cellhypoxia inducible factorhigh mobility group box
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在東南亞國家及台灣造成多次大流行的人類腸病毒71型(EV71)除了引發手足口病之外,也造成嚴重的神經學後遺症及致死性的肺水腫。肺水腫過去被認為導因於中樞神經感染後引起的自律神經系統失調。然而,局部性或全身性的促發炎因子因感染後被大量起動亦被懷疑是造成肺水腫的病理機轉之一。巨噬細胞移動抑制因子(MIF)調節發炎免疫反應及影響細胞生長調控,與許多疾病發生有關。本研究探討EV71感染橫紋肌肉瘤細胞及人類肺腺癌細胞後細胞之變化以及發炎促進物質MIF於細胞之表現。其結果發現,比起其他具臨床重症症狀的患者,在併發肺水腫患者血清中有較高的MIF被偵測到,於肺泡的洗出液則有更高的MIF偵測量。在被EV71感染後,人類肺腺癌細胞有MIF之表現。橫紋肌肉瘤細胞於感染後會發生細胞死亡現象,並釋放出高遷移率族蛋白(HMGB-1)。被釋放出的HMGB-1會誘發一連串的發炎反應。相反的,在而人類肺腺癌細胞被EV71感染後,細胞則發生細胞增生的現象。這兩種細胞的不同反應是導因於病毒的直接影響亦或受MIF之表現之影響仍待進一步之探討。另外,在人類肺腺癌細胞的研究中,缺氧誘發因子(HIF)及VEGF皆有表現量的增加。HIF與VEGF曾被發表與肺部血管滲漏進而導致肺水腫之發生有關,但本實驗的發現是否於此機制有關仍待進一步釐清。另外,本研究發現MIF於細胞的表現量與病毒感染劑量亦呈現正相關。
Human enterovirus 71 (EV71) has been the cause of several of outbreaks in countries of South-East Asia, Taiwan and China. Encephalitis and pulmonary edema are major causes of death among those who are infected by EV71. Other than neurogenic etiology, it is speculated that systemic and local pro-inflammatory responses are involved. Macrophage migration inhibitory factor (MIF) mediates immune and inflammatory reactions and influences cell growth. With rare research about relationship between MIF and EV71 infection, this study was conducted. In this study, we found A549 cells express MIF after infection. The results showed EV71-infected Rhabdomyosarcoma (RD) cells underwent cell death, releasing High mobility group 1 (HMGB-1), which is thought to induce pro-inflammatory reaction. Instead of cell death, infected human lung adenocarcinoma (A549) cells undergo proliferation, without releasing HMGB-1. Besides, Hypoxia-inducible factors (HIFs) and VEGF expression increase after infection of A549 cells. Though, it needs more research to clarify whether the previous proposed ROS-HIF-VEGF pathway, resulting in pulmonary edema, is true for this case. Furthermore, this study also revealed a positive correlation between expression level of MIF and EV71 MOI (multiplicity of infection) in A549 cells, implying MIF being a possible biomarker for assessing the severity of EV71 infection.

Contents I
Figures III
Appendix III
Chapter 1 Specific aims 1
1-1 Specific Aim 1 1
1-2 Specific Aim 2 1
Chapter 2 Background 1
2-1 Human Enterovirus 71 (EV71) 1
2-1-1 Characterization of EV71 1
2-1-2 Epidemiology of EV71 1
2-1-3 Clinical manifestations of EV71 infection 2
2-1-4 Pathogenesis of EV71 3
2-2 MIF (Macrophage migration inhibitory factors) 4
2-2-1 Characteristics of MIF 4
2-2-2 Relation of MIF with diseases 6
2-3 Hypoxia-inducible factor (HIF) 7
2-3-1 Property of HIF 7
2-3-2 Clinical significance of HIF 8
2-4 High mobility group box 1 (HMGB-1) and necrosis 9
2-5 Vascular endothelial growth factors (VEGFs) 10
2-6 Apoptosis 10
Chapter 3 Motivation and Strategies for Research 12
3-1 Specific Aim 1 12
3-2 Specific Aim 2 12
Chapter 4 Materials and Methods 14
4-1 Preparation of virus and virus titration 14
4-2 Plague assay 14
4-3 EV71 infection of cells 15
4-4 Reagents 15
4-5 Others 15
4-6 Statistical Analysis 17
Chapter 5 Results 18
5-1 The effect of EV71 infection on RD cells 18
5-2 The effect of EV71 infection on A549 cells 19
Chapter 6 Discussion 22
References 29

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