小干扰RNA对丙型肝炎病毒核心蛋白基因表达的抑制作用

发布时间：2018-04-13 13:25

本文选题：丙型肝炎病毒 + 核心蛋白　；参考：《第二军医大学》2006年硕士论文

【摘要】：丙型肝炎病毒(HCV)属黄病毒属,是导致慢性肝炎及肝硬化的主要因素。HCV基因组是9.6kb的单正链RNA,包含一个位于5′和3′非编码区之间的阅读框ORF,ORF编码全长3,008-3,037个氨基酸的前体蛋白,通过细胞及病毒的蛋白酶切割产生结构蛋白和非结构蛋白,经翻译后修饰产生至少十种不同的蛋白:核心蛋白、E1、E2、p7、非结构蛋白NS2、NS3、NS4A、NS4B、NS5A和NS5B。大多数慢性感染病人通过干扰素和利巴韦林联合治疗。因为缺少HCV颗粒的体外培养体系,对HCV这种高致病型病毒的生命周期、抗病毒及HCV疫苗的研究一直无法深入。从肝癌病人血清中克隆的亚基因组复制子在体外可以高效的复制,但不能产生病毒颗粒。最近有文献报道,建立的HCV 2a亚型的全基因组复制子JFH1在体外肝癌细胞中可高效复制,并有HCV病毒颗粒的产生,这种病毒颗粒的密度为1.15-1.17g/ml,直径为55nm,其复制可被α干扰素及多种HCV特异性抗病毒药物抑制,其感染性可以被CD81特异性抗体、包膜蛋白E2抗体及来自HCV慢性感染病人的免疫球蛋白阻断。这种病毒颗粒可以感染黑猩猩,这种培养系统为研究病毒的生命周期、宿主-病毒的相互作用、抗病毒药物及疫苗的研究提供了一个强有力的工具,是HCV研究的新突破。 HCV核心蛋白是由开放阅读框的5′端区域编码,由碱性的氨基端RNA结合区域和高度疏水的羧基端组成。成熟的21 kDa的核心蛋白由蛋白前体在191和192位氨基酸处切割产生,其19和16 kDa两种形式的蛋白也有了进一步研究。核心蛋白位于内质网的细胞质一侧,但是在细胞核中也可以检测到。核心蛋白除了对HCV RNA病毒的包装作用之外,最近研究表明,对宿主细胞的生长、凋亡及癌变也有很重要的作用,HCV核心蛋白的转化功能在老鼠胚胎成纤维细胞及转基因鼠中得到了证实。但核心蛋白影响细胞功能的机制还有待于进一步研究。细胞的信号转导异常可揭示人类疾病发生的本质,一些疾病的致病机制即源于其蛋白所制宿主细胞信号转导的紊乱。分裂原激活的蛋白激酶(mitogen-activated protein kinase, MAPK)家族属丝氨酸/苏氨酸蛋白激酶,包括MAPK/ERK(extracellular regulated protein, ERK)、JNK(C-Jun N-terminal Kinase)、ERK5/BMK1(big mito-gen-activated protein kinase 1, BMK1)及p38四个亚家族,MAPK信号途径在介导细胞对细胞内、外刺激的反应性上具有重要功能,不同亚家族的激活可产生不同生物效应:促细胞增殖、分化、凋亡等,但当其活力失控时会导致肿瘤。MAPK/ERK主要调节细胞的生长增殖,核心蛋白的表达可导
[Abstract]:Hepatitis C virus (HCV) is a major cause of chronic hepatitis and cirrhosis. The HCV genome is a single-stranded RNAs of 9.6kb that contain a reading frame ORF ORF that encodes a full length of 3008-3037 amino acids between the 5 'and 3' noncoding regions.Structural and non-structural proteins were produced by protease cleavage of cells and viruses, and modified with translation to produce at least ten different proteins: core protein E1C E2P7, nonstructural protein NS2NS3, NS4ANS4NS4BN5A and NS5B.Most patients with chronic infection are treated with interferon and ribavirin.Due to the lack of in vitro culture system of HCV particles, the research on the life cycle, antivirus and HCV vaccine of HCV, a highly pathogenic virus, has been unable to be further studied.Subgenomic replicators cloned from sera of patients with liver cancer can replicate efficiently in vitro, but can not produce viral particles.Recently, it has been reported that the whole genome replicator JFH1 of HCV 2a subtype can be highly duplicated in hepatoma cells in vitro, and HCV virus particles are produced.This viral particle has a density of 1.15-1.17g / ml and a diameter of 55nm, and its replication can be inhibited by interferon alpha and a variety of HCV specific antiviral drugs, and its infectivity can be inhibited by CD81 specific antibodies.Envelope protein E2 antibody and immunoglobulin blocking from patients with chronic HCV infection.The virus particles can infect chimpanzees. This culture system provides a powerful tool for the study of virus life cycle, host-virus interaction, antiviral drugs and vaccines, and is a new breakthrough in HCV research.The HCV core protein is encoded by the 5 'terminal region of the open reading frame and consists of an alkaline amino terminal RNA binding region and a highly hydrophobic carboxyl terminal.The mature 21 kDa core protein was produced by cleavage of the protein precursor at the 191-192 amino acids, and its 19 and 16 kDa proteins were further studied.The core protein is located on the cytoplasmic side of the endoplasmic reticulum, but can also be detected in the nucleus.In addition to packaging the HCV RNA virus, recent studies have shown that the core protein is responsible for the growth of host cells.Apoptosis and carcinogenesis also play an important role in the transformation of HCV core protein in mouse embryonic fibroblasts and transgenic mice.However, the mechanism of core protein affecting cell function remains to be further studied.The abnormal signal transduction of cells can reveal the nature of human disease. The pathogenetic mechanism of some diseases is due to the disorder of signal transduction in host cells produced by their proteins.Mitogen-activated protein kinases (MAPKs) belong to serine / threonine protein kinases, including MAPK/ERK(extracellular regulated protein, ERK5 / BMK1big mito-gen-activated protein kinase 1 (BMK1) and p38 subfamilies.The activation of different subfamilies can produce different biological effects, such as promoting cell proliferation, differentiation, apoptosis, etc., but when its activity is out of control, MAPK / ERK will mainly regulate the growth and proliferation of cells.The expression of core protein can be guided
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R373

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