The Study Of The Invasion In Vitro Of Human Ovarian Cancer HO-8910 Modified By Ezrin SiRNA

Objective:To study the effects of EZRIN SiRNA to human ovarian cancer cell line HO-8910, and to bring a new method of gene therapy for ovarian cancer.Methods:1. The structure of EZRIN SiRNA expression vector.2. Cultivated the Human ovarian cancer HO-8910 cells in vitro: cultivated HO-8910 cells in 37℃, 5%CO2, with RPMI-1640 complete medium Washed with PBS buffer, 0.25% trypsin digestion and passage.3. Transfected the EZRIN SiRNA and negative control SiRNA into HO-8910 cells by LipofectamineTM2000, using RT-PCR and Western blot to detect EZRIN gene and protein expression change, verifying the siRNA inhibition effect.4. In vitro experiments: the biology effect of EZRIN on HO-8910 cell was analyzed through the methyl thiazolyl tetrazolium test, Transwell chamber cell invasion assay in vitro.Results:1. PGPU6/GFP/Neo-siRNA-EZRIN transfected with liposome to HO-8910 cells could significantly inhibit the expression of genes and proteins of EZRIN.2. After 18 hours from the transfaction of PGPU6/GFP/siRNA-EZRIN with the liposome, HO-8910 cells began to appear green fluorescence. After 36 hours the rate of transfection could reach 60%.3. In vitro invasion assay showed that invasion cell number in recombinant plasmid group(46.7±3.1) was significant difference of empty plasmid group(91.2±1.3) and blank contrast group(91.4±1.3, P<0.05).Conclusion:1. Human ovarian cancer HO-8910 cells are successfully modified by PGPU6/GFP/Neo-siRNA-EZRIN through liposome.2. EZRIN SiRNA modified human ovarian cancer HO-8910 cells can significantly inhibit ovarian cancer HO-8910 cells in vitro invasion. As a new valuable tool,it promises to have application in the gene treatment of ovarian cancer.