A non-tight junction function of claudin-7-Interaction with integrin signaling in suppressing lung cancer cell proliferation and detachment.
A non-tight junction function of claudin-7-Interaction with integrin signaling in suppressing lung cancer cell proliferation and detachment.
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DOI：
10.1186/s12943-015-0387-0
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发表时间：
2015-06-17
期刊：
影响因子：
37.3
通讯作者：
Chen YH
中科院分区：
文献类型：
--
作者：
Lu Z;Kim DH;Fan J;Lu Q;Verbanac K;Ding L;Renegar R;Chen YH
Claudins 是紧密连接 (TJ) 膜蛋白家族，与包括癌症在内的多种人类疾病有关。 Claudin-7 是一种独特的 TJ 膜蛋白，在上皮细胞和组织中具有很强的基底外侧膜分布。因此，本研究旨在探讨claudin-7这种非TJ定位在人肺癌细胞中的功能意义。 Claudin-7 表达被慢病毒 shRNA 或靶向基因删除抑制或删除。分别采用细胞周期分析和抗体封闭方法来测定细胞增殖和细胞附着。采用电子显微镜和上皮电阻测量来检查 TJ 超微结构和屏障功能。使用免疫共定位和免疫共沉淀研究claudin-7 与整合素β1 的相互作用。使用无胸腺裸鼠分析体内肿瘤生长。 Claudin-7 与整合素 β1 共定位并形成稳定的复合物。在人肺癌细胞 (KD 细胞) 中通过慢病毒 shRNA 抑制claudin-7 表达，以及在小鼠肺部删除claudin-7 都会导致整合素 β1 和磷酸-FAK 水平降低。抑制claudin-7 表达可促进细胞生长和细胞周期进程。更重要的是，claudin-7 KD 细胞在细胞-基质相互作用方面存在严重缺陷，并且与培养板的粘附性较差，整合素 β1 表达显着降低。当在未涂覆的玻璃盖玻片上培养时，claudin-7 KD 细胞在彼此之上生长并形成球体，而对照细胞粘附良好并以单层形式生长。重新引入claudin-7 可减少细胞增殖、上调整合素β1 表达并增加细胞-基质粘附。整合素β1转染部分挽救了细胞附着缺陷。当接种到裸鼠体内时，claudin-7 KD 细胞产生的肿瘤明显大于对照细胞。在这项研究中，我们发现了claudin-7在通过整合素β1调节细胞增殖和维持上皮细胞附着方面以前未被认识的功能。本文的在线版本 (doi:10.1186/s12943-015-0387-0) 包含补充材料，可供授权用户使用。
Claudins are a family of tight junction (TJ) membrane proteins involved in a broad spectrum of human diseases including cancer. Claudin-7 is a unique TJ membrane protein in that it has a strong basolateral membrane distribution in epithelial cells and in tissues. Therefore, this study aims to investigate the functional significance of this non-TJ localization of claudin-7 in human lung cancer cells. Claudin-7 expression was suppressed or deleted by lentivirus shRNA or by targeted-gene deletion. Cell cycle analysis and antibody blocking methods were employed to assay cell proliferation and cell attachment, respectively. Electron microscopy and transepthelial electrical resistance measurement were performed to examine the TJ ultrastructure and barrier function. Co-immunolocalization and co-immunoprecipitation was used to study claudin-7 interaction with integrin β1. Tumor growth in vivo were analyzed using athymic nude mice. Claudin-7 co-localizes and forms a stable complex with integrin β1. Both suppressing claudin-7 expression by lentivirus shRNA in human lung cancer cells (KD cells) and deletion of claudin-7 in mouse lungs lead to the reduction in integrin β1 and phospho-FAK levels. Suppressing claudin-7 expression increases cell growth and cell cycle progression. More significantly, claudin-7 KD cells have severe defects in cell-matrix interactions and adhere poorly to culture plates with a remarkably reduced integrin β1 expression. When cultured on uncoated glass coverslips, claudin-7 KD cells grow on top of each other and form spheroids while the control cells adhere well and grow as a monolayer. Reintroducing claudin-7 reduces cell proliferation, upregulates integrin β1 expression and increases cell-matrix adhesion. Integrin β1 transfection partially rescues the cell attachment defect. When inoculated into nude mice, claudin-7 KD cells produced significantly larger tumors than control cells. In this study, we identified a previously unrecognized function of claudin-7 in regulating cell proliferation and maintaining epithelial cell attachment through engaging integrin β1. The online version of this article (doi:10.1186/s12943-015-0387-0) contains supplementary material, which is available to authorized users.
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影响因子：
4.8
作者：
Keely, Simon;Glover, Louise E.;Colgan, Sean P.
通讯作者：
Colgan, Sean P.
DOI：
10.1016/s0165-3806(98)00091-1
发表时间：
1998-09-10
期刊：
DEVELOPMENTAL BRAIN RESEARCH
影响因子：
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影响因子：
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作者：
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通讯作者：
Siegel, P. M.
影响因子：
8
作者：
通讯作者：
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影响因子：
5.3
作者：
Moldvay, Judit;Jaeckel, Marta;Kiss, Andras
通讯作者：
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