Inhibition of invasive and chemotactic abilities of SKOV3 cells bv human epithelial growth receptor-2 small interfering RNA / 中华妇产科杂志

ABSTRACT

Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P