RNA干扰沉默基质金属蛋白酶2基因对卵巢癌OVCAR-3细胞诱导体外血管形成能力的影响

Effect on angiogenesis induced by ovarian cancer OVCAR-3 cells in vitro when RNA interference on expression of matrix metalloproteinase-2 gene

目的探讨沉默基质金属蛋白酶2（MMP-2）基因对卵巢癌OVCAR．3细胞血管内皮生长因子（VEGF）表达的影响，以及对细胞诱导的体外血管形成能力的影响。方法合成特异性靶向基因的小干扰RNA（siRNA）并转染卵巢癌OVCAR-3细胞（MMP-2沉默组），以非特异性序列转染细胞作为阴性对照组，以培养液代替转染siRNA的试剂作为空白对照组；采用实时荧光定量PCR和Westernblot法分别检测转染后48hMMP-2及VEGF的mRNA和蛋白表达水平，通过体外血管形成实验检测卵巢癌OVCAR-3细胞诱导的体外血管形成能力。结果与阴性对照组相比，MMP-2沉默组转染48h，OVCAR-3细胞MMP．2和VEGFmRNA表达量分别下降了78．8％和75．5％（P〈0．05），蛋白表达量分别下降了81．2％和783％（P〈0．05）；体外血管形成实验显示，沉默MMP-2基因后卵巢癌OVCAR-3细胞诱导的体外血管形成能力明显降低（P〈0．05）。结论沉默MMP-2基因可抑制卵巢癌细胞VEGF表达，并能够抑制卵巢癌细胞诱导的体外血管形成能力，抑制MMP-2基因有助于抗血管形成，可能成为卵巢癌基因治疗的新靶点。

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.