Lin Yang, Cui Manhua, Xu Tianmin, Yu Wei, Zhang Lihui
Department of Gynaecology and Obstetrics, The Second Hospital of Jilin University, Changchun, Jilin 130041, P.R. China.
Mol Med Rep. 2014 Jun;9(6):2499-504. doi: 10.3892/mmr.2014.2131. Epub 2014 Apr 9.
The present study aimed to investigate the effect of downregulating cyclooxygenase‑2 (COX‑2) expression on the growth of human ovarian cancer cells. The COX‑2‑specific small interfering RNA (siRNA) plasmid vector was constructed and then transfected into ovarian cancer cells. The expression of COX‑2 mRNA and protein was detected by quantitative polymerase chain reaction and western blot analysis, respectively. Cell proliferation, apoptosis, cell cycle distribution and cell migration were assessed following knockdown of COX‑2 by RNA interference (RNAi). Western blot analysis was used to identify differentially expressed angiogenesis- and cell cycle‑associated proteins in cells with silenced COX‑2. The expression levels of COX‑2 in ovarian cancer cells transfected with siRNA were decreased, leading to a significant inhibition of ovarian cancer cell proliferation, migration and invasion. Western blot analysis revealed that silencing of COX‑2 may inhibit vascular endothelial growth factor, matrix metalloproteinase (MMP)‑2 and MMP‑9 protein expression. In conclusion, the present study demonstrated that RNAi can effectively silence COX‑2 gene expression and inhibit the growth of ovarian cancer cells, which indicates that there is a potential of targeting COX‑2 as a novel gene therapy approach for the treatment of ovarian cancer.
本研究旨在探讨下调环氧化酶-2(COX-2)表达对人卵巢癌细胞生长的影响。构建了COX-2特异性小干扰RNA(siRNA)质粒载体,然后将其转染到卵巢癌细胞中。分别通过定量聚合酶链反应和蛋白质印迹分析检测COX-2 mRNA和蛋白的表达。在通过RNA干扰(RNAi)敲低COX-2后,评估细胞增殖、凋亡、细胞周期分布和细胞迁移情况。采用蛋白质印迹分析鉴定COX-2沉默细胞中差异表达的血管生成和细胞周期相关蛋白。转染siRNA的卵巢癌细胞中COX-2的表达水平降低,导致卵巢癌细胞增殖、迁移和侵袭受到显著抑制。蛋白质印迹分析显示,COX-2沉默可能抑制血管内皮生长因子、基质金属蛋白酶(MMP)-2和MMP-9蛋白表达。总之,本研究表明RNAi可有效沉默COX-2基因表达并抑制卵巢癌细胞生长,这表明靶向COX-2作为一种新型基因治疗方法治疗卵巢癌具有潜力。