CAJ | 학술논문

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Western blot法分别检测转染后48 h MMP-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48 h,OVCAR-3细胞MMP-2和VEGF mRNA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78.3％(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.
목적 탐토침묵기질금속단백매2(MMP-2)기인대란소암OVCAR-3세포혈관내피생장인자(VEGF)표체적영향,이급대세포유도적체외혈관형성능력적영향.방법 합성특이성파향기인적소간우RNA (siRNA)병전염란소암OVCAR-3세포(MMP-2침묵조),이비특이성서렬전염세포작위음성대조조,이배양액대체전염siRNA적시제작위공백대조조；채용실시형광정량PCR화Western blot법분별검측전염후48 h MMP-2급VEGF적mRNA화단백표체수평,통과체외혈관형성실험검측란소암OVCAR-3세포유도적체외혈관형성능력.결과 여음성대조조상비,MMP-2침묵조전염48 h,OVCAR-3세포MMP-2화VEGF mRNA표체량분별하강료78.8％화75.5％(P＜0.05),단백표체량분별하강료81.2％화78.3％(P＜0.05)；체외혈관형성실험현시,침묵MMP-2기인후란소암OVCAR-3세포유도적체외혈관형성능력명현강저(P＜0.05).결론 침묵MMP-2기인가억제란소암세포VEGF표체,병능구억제란소암세포유도적체외혈관형성능력,억제MMP-2기인유조우항혈관형성,가능성위란소암기인치료적신파점.
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.