Abstract:
To construct shRNA eukaryotic expression vectors that targeting human CC chemokine receptor 5(CCR5)and to investigate the biological function of CCR5 in breast cancer cell adhesion and migration, the gene expression of CCR5 in MDA-MB-231, MDA-MB-468, T47D human breast cancer cell lines was measured by quantitative PCR, and MDA-MB-231 cells highly expressing CCR5 were used in the next experiments. Secondly, to construct shRNA eukaryotic expression vectors that express shRNA targeting human CCR5, small fragments of siRNA were designed and cloned into the Ambion shRNA vector pSilencer 2. 1-U6. The recombinant plasmids(shCCR5-1 and shCCR5-2)were then transiently transfected in MDA-MB-231 cells and CCR5 gene and protein expression were measured by quantitative PCR and Sestern blot. Thirdly, the effect of knockdown of CCR5 on MDA-MB-231 cell adhesion and migration was determined by adhesion assay and transwell chamber test, respectively. It was found that silencing of CCR5 greatly inhibited MDA-MB-231 cell adhesion and migration.