紧密连接蛋白claudin-1对HCVcc易感性的影响

目的利用HCV体外培养细胞模型,观察紧密连接蛋白claudin-1对于HCV易感性的影响。方法构建pEGFP-claudin-1真核表达载体,脂质体转染293T细胞,观察claudin-1的表达,检测claudin-1对HCVcc感染性的影响;使用化学合成claudin-1 siRNA序列,抑制Huh-7.5细胞中claudin-1的表达,检测HCVcc感染性的变化。结果 293T细胞上外源性瞬时表达claudin-1后,获得对HCVcc的易感性,化学合成的siRNA序列可有效抑制Huh-7.5细胞上的claudin-1的表达,阻断HCVcc对Huh-7.5细胞的感染。结论紧密连接蛋白clau...

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Published in临床肝胆病杂志 Vol. 28; no. 8; pp. 612 - 615
Main Author 魏欣 贾战生 王涛 李彧 马力 连建奇 王九平 张野 白雪帆 郝春秋
Format Journal Article
LanguageChinese
Published 第四军医大学唐都医院传染科,西安,710038%解放军总后勤部司令部管理保障局第一门诊部保健科,北京,100842%陕西省人民医院传染科,西安,710065 2012
Subjects
claudin-1
易感性
肝炎病毒属
连接蛋白类
易感性
连接蛋白类
肝炎病毒属
claudin -1
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ISSN1001-5256

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Summary:目的利用HCV体外培养细胞模型,观察紧密连接蛋白claudin-1对于HCV易感性的影响。方法构建pEGFP-claudin-1真核表达载体,脂质体转染293T细胞,观察claudin-1的表达,检测claudin-1对HCVcc感染性的影响;使用化学合成claudin-1 siRNA序列,抑制Huh-7.5细胞中claudin-1的表达,检测HCVcc感染性的变化。结果 293T细胞上外源性瞬时表达claudin-1后,获得对HCVcc的易感性,化学合成的siRNA序列可有效抑制Huh-7.5细胞上的claudin-1的表达,阻断HCVcc对Huh-7.5细胞的感染。结论紧密连接蛋白claudin-1影响细胞对HCV的易感性,可能与HCV的嗜肝性有关。
Bibliography:Objective To study the influence of the tight junction protein, claudin - 1, on susceptibility to HCV in cell culture （HCVcc）. Methods An enhanced green fluorescent protein （ EGFP eukaryotie expression vector, pEGFP - claudin - 1, was constructed and transfect- ed into human embryonic kidney （HEK） -293T cells with lipofectamine. The expression of claudin - 1 and corresponding susceptibility to HCVcc was analyzed by fluorescence assay and RT -PCR, respectively. A chemically synthesized small interfering （si）RNA targeting clan- din - 1 was transfeeted into Huh - 7.5 hepatoma cells and the effect of claudin - 1 silencing on susceptibility to HCVcc was tested by West- em blotting and quantitative RT - PCR. Results Expression of elaudin - 1 in HEK - 293T cells conferred susceptibility to HCV infection. Ctandin - 1 expression was reduced in Huh -7.5 cells by targeted siRNA. Susceptibility to HCVcc was markedly reduced in claudin - 1 - HCV silenced Huh - 7. 5 cells. Conclusion Tight junction protein claudin - 1 expr
ISSN:1001-5256