| 摘要: |
| 目的:探讨基质细胞衍生因子1(SDF1)及其受体CXC趋化因子受体4(CXCR4)在肾癌细胞转移中的作用机制,观察不同区段CXCR4在肾癌细胞内的定位。方法:应用核定位分析软件结合实验,构建不同长度的CXCR4与绿色荧光蛋白pEGFPN1重组表达载体,转染肾透明细胞癌A498细胞株后用共聚焦显微镜观察重组表达载体在细胞内的定位。结果:生物信息学分析软件PSORT Ⅱ Prediction发现第146～149位氨基酸残基RPRK可能是CXCR4的核定位序列,我们构建的重组质粒EGFPCXCR4(1～510 bp)、EGFPCXCR4(1～765 bp)、野生型全长EGFPCXCR4分别加SDF1刺激因子后其表达产物主要呈细胞核分布,而加入SDF1刺激因子后重组质粒EGFPCXCR4(1～267 bp)的表达产物呈细胞质分布,未经SDF1刺激野生型全长EGFPCXCR4其表达产物呈细胞质分布,与生物信息学分析软件预测结果初步吻合。结论:CXCR4的第90～170位氨基酸残基含有核定位序列,为进一步精确定位CXCR4在肾癌细胞内的核定位序列以及寻找抑制肾癌转移的可能靶标奠定了实验和理论基础。 |
| 关键词: 肾肿瘤 趋化因子 核定位序列 亚细胞定位 |
| DOI：10.3724/SP.J.1008.2009.0990 |
| 投稿时间：2009-03-17修订日期：2009-05-07 |
| 基金项目:国家自然科学基金(30873040/C171010). |
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| Preliminary study on CXCR4 nuclear localization sequence in metastatic renal cell carcinoma |
| LIU Qian,WANG Lin-hui,YANG Qing,XU Bin,SUN Ying-hao* |
| (Department of Urology,Changhai Hospital,Second Military Medical University,Shanghai 200433,China) |
| Abstract: |
| Objective:To investigate the role of SDF1/CXCR4 in metastasis of renal cell carcinoma and to observe the intracellular location of different CXCR4 segments in renal carcinoma cells.Methods: The potential nuclear localization sequences of different CXCR4 were discovered by nuclear localization software and experiments.Full length and truncated forms of CXCR4 were fused with green fluorescent protein pEGFPN1 and their influence on subcellular localization was examined by confocal microscopy after transfecting them into renal carcinoma cell line A498.Results: Analysis with PSORT Ⅱ Prediction revealed that the nuclear localization sequence region of CXCR4 was located between amino acids 146 and 149(RPRK).Expression products of the recombinant plasmids with SDF1 stimulation,including EGFPCXCR4(1510 bp),EGFPCXCR4(1765 bp) and wildtype EGFPCXCR4,were mainly located in the cell nuclei.However,expression product of EGFPCXCR4(1267 bp) with SDF1 stimulation was mainly located in the renal cell cytoplasm.Expression product of wildtype EGFPCXCR4 full length plasmid without SDF1 stimulation was mainly located in the cell cytoplasm; these results accorded with the results of bioinformatics analysis.Conclusion: Nuclear localization sequence of CXCR4 is located in the amino acids 90 to 170,which provides a theoretical basis for further clarifying the nuclear localization sequences of CXCR4 in renal cell carcinoma cells and for finding new potential target for inhibiting the metastasis of renal cell carcinoma. |
| Key words: kidney neoplasms chemokine nuclear localization sequence subcellular localization |
