ABSTRACT
Objective To investigate expression of VEGF and the in vitro angiogenesis ability induced by ovarian cancer cells after RNA interference on expression of matrix metalloproteinase-2 (MMP-2)gene.Methods One specific target sequence of MMP-2 and one non-specific sequence (NC group) were chosen,the DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells,mRNA and protein expression of MMP-2 and VEGF genes were examined by RT-PCR and Western blot analysis,and the angiogenesis ability was detected by in vitro angiogenic assay.Results When compared with the NC group,the mRNA expression of MMP-2 and VEGF were decreased by 78.8 ％ and 75.5 ％ (P ＜ 0.05) in 48 h after transfected,respectively,and protein expression was decreased by 81.2 ％ and 78.3 ％ (P ＜ 0.05) at the same time point.In vitro angiogenic assay suggested that the ability of angiogenesis was inhibited when down-regulated of MMP-2 gene (P ＜ 0.05).Conclusion Down-regulation of MMP-2 gene in ovarian cancer cells by RNA interference could inhibit its VEGF expression and in vitro angiogenesis induced by ovarian cancer cells,which suggestes that the inhibition of MMP-2 gene has an anti-angiogenesis effect,and MMP-2 gene could be a potential target for ovarian cancer gene-therapy.
ABSTRACT
Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 ％,78.8 ％ and 78.4 ％(P＜ 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 ％,81.2 ％ and 76.4 ％(P＜ 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P＞ 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 ％ at 60 min and 44.8 ％ at 90 min (P＜ 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 ％ and 35.8 ％(P＜0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies.