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质膜水通道蛋白-1活性影响HT20结肠癌细胞迁移。

Aquaporin-1 activity of plasma membrane affects HT20 colon cancer cell migration.

作者信息

Jiang Yong

机构信息

Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.

出版信息

IUBMB Life. 2009 Oct;61(10):1001-9. doi: 10.1002/iub.243.

Abstract

Recent studies revealed an important role of aquaporins (AQPs) in cell migration and migration-associated cell function such as angiogenesis, wound healing, and neutrophil motility. Migration of tumor cells is a crucial step in tumor invasion and metastasis. In the present study, we investigated the expression of AQP1 in human HT20 colon cancer cells and characterized its function in cell migration. By reverse transcription-polymerase chain reaction (RT-PCR) and immunoblot analysis, expression of AQP1 was identified in HT20 cell lines. Immunofluorescence analysis indicated expression of AQP1 protein in the plasma membrane of HT20 cells. The recombinant adenovirus expressing human AQP1 increased the mRNA and protein expression of AQP1 in HT20 cells. In contrary, the RNA interference vector of AQP1 effectively inhibited the mRNA and protein expression of AQP1 in HT20 cells. Adenovirus-mediated high expression of AQP1 in HT20 cells increased relative plasma membrane water permeability and migration rate in both wound healing and invasive transwell migration assays. In contrary, RNA interference vector-mediated low expression of AQP1 in HT20 cells reduced relative plasma membrane water permeability and migration rate. AQP1 expression induced relocalization of actin protein and activation of RhoA and Rac. In nude mice, AQP1 increased extravasation of HT20 Cells in lung after tail vein injection. The results provided the direct evidence that aquaporin-mediated plasma membrane water permeability plays an important role in colon cancer cell migration and may be associated with colon cancer invasion and metastasis.

摘要

最近的研究揭示了水通道蛋白(AQPs)在细胞迁移以及与迁移相关的细胞功能(如血管生成、伤口愈合和中性粒细胞运动)中发挥的重要作用。肿瘤细胞的迁移是肿瘤侵袭和转移的关键步骤。在本研究中,我们调查了水通道蛋白1(AQP1)在人HT20结肠癌细胞中的表达,并对其在细胞迁移中的功能进行了表征。通过逆转录-聚合酶链反应(RT-PCR)和免疫印迹分析,在HT20细胞系中鉴定出了AQP1的表达。免疫荧光分析表明HT20细胞质膜中有AQP1蛋白表达。表达人AQP1的重组腺病毒增加了HT20细胞中AQP1的mRNA和蛋白表达。相反,AQP1的RNA干扰载体有效抑制了HT20细胞中AQP1的mRNA和蛋白表达。腺病毒介导的HT20细胞中AQP1的高表达在伤口愈合和侵袭性Transwell迁移试验中均增加了相对质膜水通透性和迁移率。相反RNA干扰载体介导的HT20细胞中AQP1的低表达降低了相对质膜水通透性和迁移率。AQP1的表达诱导肌动蛋白蛋白重新定位以及RhoA和Rac激活。在裸鼠中,尾静脉注射后AQP1增加了HT20细胞在肺中的外渗。这些结果提供了直接证据,表明水通道蛋白介导的质膜水通透性在结肠癌细胞迁移中起重要作用,并且可能与结肠癌的侵袭和转移有关。

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