Preliminary study on CXCR4 nuclear localization sequence in metastatic renal cell carcinoma
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Supported by National Natural Science Foundation of China (30873040/C171010).

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    Abstract:

    Objective:To investigate the role of SDF1/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 pEGFPN1 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 SDF1 stimulation,including EGFPCXCR4(1510 bp),EGFPCXCR4(1765 bp) and wildtype EGFPCXCR4,were mainly located in the cell nuclei.However,expression product of EGFPCXCR4(1267 bp) with SDF1 stimulation was mainly located in the renal cell cytoplasm.Expression product of wildtype EGFPCXCR4 full length plasmid without SDF1 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.

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History
  • Received:March 17,2009
  • Revised:May 07,2009
  • Adopted:July 22,2009
  • Online: September 17,2009
  • Published:
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