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首页> 外文期刊>Journal of Cellular Physiology >Expression and targeting of the tight junction protein CLDN1 in CLDN1-negative human breast tumor cells.
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Expression and targeting of the tight junction protein CLDN1 in CLDN1-negative human breast tumor cells.

机译：表达和紧密连接的目标蛋白质CLDN1 CLDN1-negative人类乳房肿瘤细胞。

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摘要

Claudins and occludin constitute the major transmembrane proteins of tight junctions (TJs). We have previously identified the human homologue of the murine Cldn1, CLDN1 (SEMP1) that is expressed in normal, mammary gland-derived epithelial cells but is absent in most human breast cancer cell lines. To investigate the potential functions of CLDN1 protein in tumor and normal epithelial cells, we developed an I-NGFR retroviral vector and monoclonal anti-CLDN1 antibody. In subconfluent and confluent breast cancer cells, MDA-MB-435 and MDA-MB-361, endogenous CLDN1 expression was not detected by an anti-CLDN1 monoclonal antibody by Western blot analysis or quantitative RT-PCR. When CLDN1-negative breast cancer cell lines were transduced with a CLDN1 retrovirus the cells express CLDN1 mRNA constitutively as shown by quantitative RT-PCR. Immunofluorescence analyses of the CLDN1 retroviral transduced breast tumor cells using monoclonal antibodies against CLDN1 reveals a subcellular distribution at cell-cell contact sites similar to the CLDN1 homing pattern in T47-D cells, which express endogenous CLDN1. This cell-cell contact co-localization of CLDN1 was evident in CLDN1-transduced breast tumor cells which fail to express occludin protein (MDA-MB-361 and MDA-MB-435) and express relatively little ZO-1 protein (MDA-MB-435), suggesting that other proteins may be responsible for targeting of CLDN1 to cell-cell contact sites. The re-expression of CLDN1 decreases the paracellular flux of 3 and 40 kDa dextran despite the absence of occludin in the MDA-MB-361 tumor cells. Our findings indicate that in CLDN1-negative breast tumor cells, the basal protein partner requirements for physiological homing of the CLDN1 protein are intact, and that CLDN1 gene transfer and protein expression itself might be sufficient to exert a TJ-mediate gate function in metastatic tumor cells even in the absence of other TJ-associated proteins, such as occludin.
机译：Claudins和occludin构成主要的跨膜蛋白的紧密连接(套)。我们以前确定人类的同系物的小鼠Cldn1 Cldn1 (SEMP1)表示在正常乳腺gland-derived在大多数人类上皮细胞但缺席乳腺癌细胞系。CLDN1蛋白在肿瘤的潜在功能正常上皮细胞,我们开发了一个I-NGFR逆转录病毒载体和单克隆anti-CLDN1抗体癌细胞,mda - mb - 435和mda - mb - 361,内源性CLDN1表达式没有检测到一个anti-CLDN1单克隆抗体免疫印迹分析或者定量rt - pcr。CLDN1-negative乳腺癌细胞系转导CLDN1逆转录病毒的细胞表达CLDN1信使rna结构上如图所示定量rt - pcr。的CLDN1逆转录病毒转导乳腺肿瘤细胞对CLDN1使用单克隆抗体揭示了一个亚细胞分布信息联系网站类似于CLDN1自导模式内生CLDN1 T47-D细胞中表达。这信息联系co-localization CLDN1很明显在CLDN1-transduced乳腺肿瘤细胞不表达occludin的蛋白质(mda - mb - 361和mda - mb - 435)和表达相对较少的ZO-1蛋白质(mda - mb - 435),建议其他蛋白质可能是负责任的针对CLDN1信息联系网站。paracellular通量3和40 kDa右旋糖酐尽管没有occludin的mda - mb - 361肿瘤细胞。CLDN1-negative乳腺癌肿瘤细胞、基底蛋白质生理伙伴要求归航的CLDN1蛋白质完好无损,CLDN1基因转移和蛋白表达本身足以产生TJ-mediate门在转移性肿瘤细胞即使在函数没有其他TJ-associated蛋白质,如occludin。

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