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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10μmol/L组、柚皮苷20μmol/L组、柚皮苷40μmol/L组、阳性对照组(塞来昔布80μmol/L)。MTT法测定柚皮苷对人卵巢癌SKOV3细胞增殖的影响,运用Re-al-time PCR和Western Blot法测定培养48h后各组细胞中COX-2mRNA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48h后,Real-time PCR结果显示,与空白对照组(1.094±0.053)比较,10、20μmol/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40μmol/L柚皮苷则明显下调COX-2mRNA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,10μmol/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、40μmol/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-2mRNA和蛋白的表达。
Objective: To study the effect of naringin on mRNA and protein expression of cyclooxygenase 2 (COX-2) in human ovarian cancer cell line SKOV3. Methods: Human ovarian cancer SKOV3 cells were routinely cultured and divided into blank control group, naringin 10μmol / L group, naringin 20μmol / L group, naringin 40μmol / L group, positive control group (celecoxib 80μmol / L). MTT assay was used to determine the effect of naringin on the proliferation of human ovarian cancer cell line SKOV3. The expression of COX-2 mRNA and protein in each group was determined by Re-al-time PCR and Western Blot. Results: MTT assay showed that the naringin treatment groups inhibited the growth of human ovarian cancer SKOV3 cells in a time-dependent and dose-dependent manner. Real-time PCR results showed that 10,20μmol / L naringin inhibited the COX-2 mRNA expression in SKOV3 cells (0.828 ± 0.006, 0.753 ± 0.011, P <0.05). However, 40 μmol / L naringin down-regulated the expression of COX-2 mRNA significantly (0.412 ± 0.216, P <0.01), similar to that of celecoxib group (0.321 ± 0.017). The results of Western Blot showed that the relative expression level of 10μmol / L naringin group (1.7925 ± 0.0880) was slightly up-regulated compared with the blank control group (1.7325 ± 0.0826), but there was no significant difference between the two groups (20,40μmol / L The naringin group could significantly down-regulate the COX-2 protein expression in SKOV3 cells (1.2225 ± 0.0822,1.2725 ± 0.0763, P <0.05), and also inhibit COX-2 protein expression in celecoxib group. CONCLUSION: Naringin can inhibit the proliferation and inhibit the expression of COX-2 mRNA and protein in human ovarian cancer SKOV3 cells in vitro.