Identification of nuclear localization sequence of CXCR4 in renal cell carcinoma by constructing expression plasmids...

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.