CXCR4 gene expression promotes K562 cells chemotaxis / 基础医学与临床

ABSTRACT

Objective To construct the tranfected cell expressing the human CXCR4 gene and to identify the effect on its immigration. Methods Total RNA was isolated from peripheral blood monouclear cell (PBMC),the full-length CXCR4 gene was amplified by RT-PCR and was inserted into plasmids PBudCE4.1 which have two promtors, after the identification by digestion and sequencing,the recombinant was transfected into K562 cell by lipofectamineTM2000. After screening culture by zeocin, stable transfected K562 cell line was established, and transcription and exression of CXCR4 were checked by flow cytometry; the chemotactic activity of K562 cell transfected and untrandfected CXCR4 was analyzed by Transell plate. Results The eukaryotic expression plasmid PBudCE4.1/ CXCR4 was constructed successfully. The stable trasfected K562/CXCR4 cell lines which highly express CXCR4 was established,the chemotactic activity of K562/CXCR4 was increased significiantly than K562. ConclusionCXCR4 transfected K562 cell line was successfully established, and it can make the basis for the further research on mechanism of extramedullary infiltration in leukemia.