摘要
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.
出处
《肿瘤研究与临床》
CAS
2012年第12期793-796,共4页
Cancer Research and Clinic
基金
广东省科技厅项目(20098060700080)
广州医学院课题(2011A16)