柚皮苷对人卵巢癌SKOV3细胞环氧化酶2 mRNA及蛋白表达水平的影响

目的: 研究中药柚皮苷对人卵巢癌SKOV3细胞环氧化酶2(COX-2)mRNA及蛋白表达水平的影响。方法: 常规培养人卵巢癌SKOV3细胞,分为空白对照组、柚皮苷 10 μmol/L 组、柚皮苷 20 μmol/L 组、柚皮苷 40 μmol/L 组、阳性对照组(塞来昔布 80 μmol/L)。MTT法测定柚皮苷对人卵巢癌SKOV3细胞增殖的影响,运用Real-time PCR和Western Blot法测定培养48 h后各组细胞中COX-2 mRNA和蛋白表达水平的变化。结果: MTT检测法显示,柚皮苷各浓度处理组,时间依赖性和剂量依赖性地抑制人卵巢癌SKOV3细胞生长。经药物处理48 h后,Real-time PCR结果显示,与空白对照组(1.094±0.053)比较,10、20 μmol/L 柚皮苷即对SKOV3细胞COX-2 mRNA表达抑制作用(0.828±0.006,0.753±0.011,P<0.05),而 40 μmol/L 柚皮苷则明显下调 COX-2 mRNA的表达(0.412±0.216,P<0.01),与塞来昔布组(0.321±0.017)的抑制程度相近。Western Blot法检测结果显示,与空白对照组(1.7325±0.0826)相比,10 μmol/L 柚皮苷组相对表达量(1.7925±0.0880)虽略有上调,但无统计学差异,20、40 μmol/L 柚皮苷组均可明显下调SKOV3细胞COX-2蛋白的表达(1.2225±0.0822,1.2725±0.0763,P<0.05),塞来昔布组也明显抑制COX-2蛋白的表达。结论: 柚皮苷可抑制人卵巢癌SKOV3细胞体外增殖并抑制COX-2 mRNA和蛋白的表达。

Effects of Naringin on expression of COX-2 mRNA and protein in Human ovarian cancer cell line SKOV3

AIM: To investigate effects of naringin on expression of COX-2 mRNA and protein of Human ovarian cancer cell line SKOV3.METHODS: SKOV3 cells were cultured in vitro and divided into five groups randomly, namely control group, naringin group (10 μmol/L), naringin group (20 μmol/L), naringin group (40 μmol/L) and celecoxib group (80 μmol/L). The inhibition effects of naringin on cell- proliferation were determined by MTT method. The expression of COX-2 mRNA and protein were evaluated by Western Blot and Real-time quantitative PCR after forty-eight hours.RESULTS: Naringin could inhibit the growth of SKOV3 cells, and with the increase of the dosage and time, the tumor growth inhibition rate increased significantly (P<0.01).Compared with control group,the expression of COX-2 mRNA was down-regulated significantly by every dosage of naringin and celecoxib (0.828±0.006, 0.753±0.011, 0.412±0.216, 0.321±0.017). Meanwhile, the expression of COX-2 protein was down-regulated significantly by every dosage of naringin and celecoxib (1.7925±0.088, 1.2225±0.0822, 1.2725±0.07632, 0.3075±0.0977).CONCLUSION: The appropriate dosage of naringin can inhibit SKOV3 cells growth in vitro, and can down-regulate the expression of COX-2 mRNA and protein.