Identification of nuclear localization sequence of CXCR4 in renal cell carcinoma by constructing expression plasmids of different deletants
Identification of nuclear localization sequence of CXCR4 in renal cell carcinoma by constructing expression plasmids of different deletants
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构建不同缺失子表达质粒鉴定肾细胞癌CXCR4核定位序列
DOI：
10.1016/j.plasmid.2009.09.004
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发表时间：
2010-01-01
期刊：
影响因子：
2.6
通讯作者：
Sun, Ying-Hao
中科院分区：
文献类型：
--
作者：
Wang, Lin-Hui;Liu, Qian;Sun, Ying-Hao
我们先前首次发现，CXCR4的核定位可能与肾细胞癌（RCC）的转移有关，并且存在一个核定位序列（NLS），它在与配体SDF - 1结合后诱导CXCR4转移至细胞核。以我们先前构建的pEGFP - CXCR4为模板，扩增相应的目标区域。然后将扩增的PCR产物进行酶切，并插入到pMD19 - T simple载体中，再亚克隆到pEGFP - N1载体。成功构建了一个包含CXCR4不同区域的重组表达载体。将重组表达载体转染到肾细胞癌A498细胞后，通过共聚焦显微镜检测重组蛋白在细胞内的定位。发现CXCR4的核定位序列位于第90 - 170位氨基酸，这与生物信息学分析软件的结果相符。本研究首次发现了CXCR4的NLS区域，这可能对寻找抑制肾细胞癌转移的新策略具有重要价值。（C）2009爱思唯尔公司。保留所有权利。
We previously firstly discovered that CXCR4 nuclear localization may be responsible for metastasis of renal cell carcinoma (RCC), and that there was a nuclear localization sequence (NLS) that induced CXCR4 to transfer to the nucleus after combining with its ligand SDF-1. Using our previously constructed pEGFP-CXCR4 as the template, corresponding objective regions were amplified. The amplified PCR products were then digested and inserted into the pMD19-T simple vector and subcloned into the pEGFP-N1 vector. A recombinant expression vector containing different regions of CXCR4 was successfully constructed. After transfecting the recombinant expression vectors to RCC A498 cells, the intracellular locations of recombinant protein were examined by confocal microscopy. It was found that nuclear localization sequence of CXCR4 was located in amino acids 90-170, which accorded with the results of bioinformatics analysis software. The present study firstly discovered the NLS region of CXCR4, which may prove valuable for seeking new strategies to inhibit metastasis of RCC. (C) 2009 Elsevier Inc. All rights reserved.