Construction of Lentiviral Vector of miR-125b and the Effect of miR-125b on Cell Proliferation and Migration of Ovarian Cancer SKOV3 Cells

Abstract: We constructd the lentiviral vector expressing miR-125b and observed the effect of miR-125b on ovarian cancer cell proliferation and migration. The PCR amplification of miR-125b precursor sequence was linked with linearized GP-Supersilencing Vector to generate the recombinant lentiviral vector expressing miR-125b. The 293T cells were transfected with the plasmids of recombinant lentiviral vector， pGag/Pol， pRev and pVSV-G plasmids to package the lentivirus. SKOV3 cells were infected by obtained lentivirus and screened with puromycin. The expression of miR-125b in SKOV3 cells was detected by Real-time qPCR and the expression of potential target gene HER-2 was tested by Western blot. Cell proliferation and cell migration were assessed using the MTT assay and transwell migration assay. The results showed that the lentiviral vector expressing miR-125b was successfully constructed and achieved the over-expression of miR-125b after infected ovarian cancer SKOV3 cells. The over-expression of miR-125b inhibited cell proliferation and migration of SKOV3 cells and decreased the expression of HER-2 gene. We concluded that miR-125b could inhibit the proliferation and migration of SKOV3 cells through down-regulation of HER-2 expression.

CSTR: 32200.14.cjcb.2014.04.0012