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Interfering CXCR4 expression inhibits proliferation, adhesion and migration of breast cancer MDA-MB-231 cells
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Abstract:
Objective: To construct short-hairpin RNA (shRNA) eukaryotic expression vector targeting CXC chemokine receptor 4 (CXCR4), and to observe its impact on the proliferation, adhesion and migration of human breast cancer MDA-MB-231 cells. Methods: The fragments of CXCR4 shRNA were synthesized and cloned into pGCsi-U6-Neo-GFP vector. The recombinant plasmids were transfected into 293T cells and the most effective interfering vector was selected. MDA-MB-231 cells were transfected by liposome assay. The effects of silencing CXCR4 expression by shRNA on the growth, adhesion and migration of MDA-MB-231 cells were determined by CCK8, cell-matrix adhesion and wound healing assays, respectively. Results: The shRNA eukaryotic expression vectors targeting CXCR4 (CXCR4-shRNA) were successfully constructed and transfected into 293T cells. RT-PCR and Western blotting results showed that the maximum inhibitory rate of CXCR4 expression was 81.3%. CXCR4-shRNA transfection significantly inhibited the proliferation of MDA-MB-231 cells (P<0.05) and the adhesion between MDA-MB-231 cells and extracellular matrix (P<0.05). Wound healing experiment showed that the migration distance of MDA-MB-231 cells in CXCR4-shRNA transfection group was significantly lower than those in the control plasmid and the blank control group (P<0.01). Conclusion: CXCR4-shRNA interfering vector can specifically inhibit CXCR4 expression, proliferation, adhesion and migration of MDA-MB-231 cells.
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Project Supported:
Project supported by the Research Foundation from Shanghai Education Commission (No.09yz79)
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