The Effects Of CXCR4Gene Silencing On Cervix Cancer Cells Migration And Invasion

Objective:To investigate the effect of chemoking stromal cell derived factor(SDF-1)/CXCR4axis on the invasion and metastasis of cervix cancer; and to construct a CXCR4specific shRNA recombinant plasmid vector and study its inhibiting effect on invasion capacity in vitro of cervix cancer.Methods:(1) A CXCR4specific recombinant plasmid vector was prepared and transfected into the Hela cell and caski cell with lipofectamine2000. RT-PCR and Western blot were used to detect the mRNA and protein expression of CXCR4, respectively.(2) Invasion capability in vitro of the cells was evaluated by Scratch test and Adhesion test.(3) Western blot were used to detect the protein expression of MMP-9and VEGF-C.Results:(1)CXCR4specific shRNA was constructed and transfected into the Hela and Caski cell line. Agarose gel electrophoresis confirmed the vector had been inserted in the production of Realtime-PCR, and sequencing result showed that composite CXCR4-shRNA inserting correctly in the positive clone. Both the CXCR4protein and mRNA expression in the CXCR4-shRNA group were significantly lower than that of the control group. The CXCR4shRNA transfection showed no signifcant effect on the distribution of cell cycle.(2) Scratch test and Adhesion test showed that the invasion capabilities of Hela and Caski cell decreased substantially(P<0.05).(3) CXCR4specific shRNA was constructed and transfected into the Hela and Caski cell line. Wester blot showed that MMP-9and VEGF-C protein expression in the CXCR4-shRNA group were significantly lower than that of the control group(p<0.05).Conclusion:(1) CXCR4make a impotant role in invasion and proliferation capacity of Hela and Caski cell in vitro.(2) CXCR4gene silencing decreased the invasion and migration capability of cervical cancer cells.the mechanism may be that blocking the SDF-1/CXCR4signaling pathway, the expression of VEGF-C and MMP-9inhibited in cervical cancer cells.