2010•Xiandai shengwu yixue jinzhanRequires access
Effect of lentiviral vector of shRNA on CXCR7 expression in human hepatoma cells
Ying Huang, Qingbo Wang, Wen Jian
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Abstract
Objective:To investigate the effect of CXCR7 short hairpin RNA(shRNA) lentiviral vector on CXCR7 expression in human hepatocellular carcinomas cell line HepG2.Methods:Four double-strand shRNAs targeting CXCR7 were designed,synthesized and cloned into the Age I and EcoRI restriction sites of pGCSIL-RFP lentiviral vector,respectively.The over-expression lentiviral vector(pEGFP-N1-3FLAG-CXCR7) containing CXCR7 mRNA was constructed,and co-transfected into HEK293T cells together with pGCSIL-RFP-CXCR7-shRNAx to investigate the optimal CXCR7-shRNA which had apparent knock-down capacity.The obtained LV-CX-CR7-shRNA lentivirus was infected into HepG2 cells and the expression of CXCR7 mRNA and protein in HepG2 cells was explored by RT-PCR and Western blot assay.Results:pGCSIL-RFP-CXCR7 shRNA2 with apparent knock-down capacity was constructed success-fully.The expression of CXCR7 mRNA and protein was down-regulated in HepG2 cells following infection with LV-CXCR7-shRNA2.Conclusion:The recombinant lentiviral vector LV-CXCR7-shRNA2 was constructed successfully and it can inhibited the expression of CXCR7 in HepG2 cells effectively.