紧密连接蛋白claudin-1对HCVcc易感性的影响

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

紧密连接蛋白claudin-1对HCVcc易感性的影响

1. 第四军医大学唐都医院传染科
2. 解放军总后勤部司令部管理保障局第一门诊部保健科
3. 陕西省人民医院传染科

基金项目:

国家自然科学基金(81101246)；陕西省自然科学基金(2011JQ4004)；

详细信息

中图分类号: R512.62
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出版历程
- 出版日期: 2012-08-20

Influence of tight junction protein claudin-1 on the susceptibility to HCVcc

Research funding:

摘要

摘要: 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。
- 连接蛋白类 /
- claudin-1 /
- 肝炎病毒属 /
- 易感性
Abstract: 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 eukaryotic expression vector, pEGFP-claudin-1, was constructed and transfected 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 claudin-1 was transfected into Huh-7.5 hepatoma cells and the effect of claudin-1 silencing on susceptibility to HCVcc was tested by Western blotting and quantitative RT-PCR. Results Expression of claudin-1 in HEK-293T cells conferred susceptibility to HCV infection.Claudin-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 expression is an important determinant of a cell's susceptibility to infection in culture and may contribute to virus tropism for particular cell phenotypes.
- connexins /
- claudin-1 /
- hepacivirus /
- susceptibility

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参考文献(12)

[1]Shepard CW, Finelli L, Alter MJ.Global epidemiology of hepati-tis C virus infection[J].Lancet Infect Dis, 2005, 5 (9) :558-567.

[2]Evans MJ, von Hahn T, Tscherne DM, et al.Claudin-1 is ahepatitis C virus co-receptor required for a late step in entry[J].Nature, 2007, 446 (7137) :801-805.

[3]马力, 魏欣, 张野, 等.体外培养的嵌合体丙型肝炎病毒感染Huh7.5细胞的透射电镜观察[J].中华微生物学和免疫学杂志, 2009, 29 (6) :568-573.

[4]魏欣, 张野, 李军, 等.真核表达载体pEGFP-claudin-1的构建及其在293T细胞中的表达[J].细胞与分子免疫学杂志, 2008, 24 (5) :444-446.

[5]Van Itallie CM, Anderson JM.Claudins and epithelial paracel-lular transport[J].Annu Rev physiol, 2006, 68:403-429.

[6]Dhawan P, Singh AB, Deane NG, et al.Claudin-1 regu-lates cellular transformation and metastatic behavior in coloncancer[J].J Clin Invest, 2005, 115 (7) :1765-1776.

[7]Roccasecca R, Ansuini H, Vitelli A, et al.Binding of the hep-atitis C virus E2 glycoprotein to CD81 is strain specific and ismodulated by a complex interplay between hypervariable re-gions 1 and 2[J].J Virol, 2003, 77 (3) :1856-1867.

[8]Elbashir SM, Harborth J, Lendeckel W, et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mamma-lian cells[J].Nature, 2001, 411 (6836) :494-498.

[9]Walters RW, Freimuth P, Moninger TO, et al.Adenovirus fi-ber disrupts CAR-mediated intercellular adhesion allowingvirus escape[J].Cell, 2002, 110 (6) :789-799.

[10]Barton ES, Forrest JC, Connolly JL, et al.Junction adhesionmolecule is a receptor for reovirus[J].Cell, 2001, 104 (3) :441-451.

[11]Reynolds GM, Harris HJ, Jennings A, et al.Hepatitis C virusreceptor expression in normal and diseased liver tissue[J].Hepatology, 2007, 47 (2) :418-427.

[12]Brimacombe CL, Grove J, Meredith LW, et al.Neutralizingantibody-resistant hepatitis C virus cell-to-cell transmis-sion[J].J Virol, 2011, 85 (1) :596-605.

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