槲皮素与FSCN1基因siRNA联合对卵巢癌生长抑制及免疫功能的影响研究

Effects of quercetin combined with FSCN1 gene siRNA on growth inhibition and immune function in ovarian cancer

目的探讨槲皮素与FSCN1基因siRNA对卵巢癌细胞活力、凋亡及免疫功能的影响及机制。方法人卵巢癌SKOV3细胞分为空白对照组、NC组(转染阴性对照siRNA)、槲皮素组(50μmol/L的槲皮素处理细胞后瞬时转染阴性对照siRNA)、si-FSCN1组(细胞转染靶向抑制FSCN1的siRNA)和槲皮素+si-FSCN1组(50μmol/L的槲皮素处理细胞后瞬时转染靶向抑制FSCN1的siRNA),siRNA转染参照Lipofectamine TM 2000说明。各组细胞处理48 h,Western blotting检测FSCN1、β-catenin、细胞周期蛋白D1(cyclin D1)、Bax和血管内皮生长因子(VEGF)的蛋白表达;MTT检测细胞活力;流式细胞术检测细胞凋亡率。结果 SKOV3细胞转染si-FSCN1后FSCN1蛋白表达明显降低,与空白对照组比较差异显著(P<0.05)。槲皮素组和si-FSCN1组细胞活力及β-catenin、cyclin D1和VEGF的蛋白表达均显著低于NC组,高于槲皮素+si-FSCN1组,细胞凋亡率及Bax蛋白表达显著高于NC组,低于槲皮素+si-FSCN1组(P<0.05)。结论槲皮素或FSCN1基因siRNA均能抑制卵巢癌细胞活力,诱导细胞凋亡,抑制免疫抑制因子VEGF表达,联合使用效果更强,机制与下调Wnt/β-catenin信号通路有关。

Objective To investigate the effects of quercetin and FSCN1 gene siRNA on viability,apoptosis and immune function of ovarian cancer cells. Methods Human ovarian cancer SKOV3 cells were divided into blank control group,NC group（transfected negative control siRNA）,quercetin group（negative control siRNA was transient transfection after cells were treated with 50 ？ mol/L quercetin）,si-FSCN1 group（FSCN1 siRNA were transfected to cells） and quercetin ＋ si-FSCN1 group（FSCN1 siRNA were transfected to cells after cells were treated with 50 ？ mol/L quercetin）,siRNA transfection referred to Lipofectamine TM 2000. The cells in each group were treated 48 h,the expression of FSCN1,β-catenin,cyclin D1,Bax and VEGF protein were detected by Western blotting. Cell viability was detected by MTT. The apoptosis rate was detected by flow cytometry. Results The expression of FSCN1 protein in KOV3 cells was significantly decreased after si-FSCN1 was transfected,which was significantly different from that in the blank control group（P〈0. 05）. The cell vitality and the expression of FSCN1,β-catenin,cyclin D1 and VEGF protein were significantly lower in quercetin group and si-FSCN1 group compared with NC group,higher compared with quercetin ＋ si-FSCN1 group,the apoptosis rate and the expression of Bax protein were significantly higher compared with NC group,lower compared with quercetin ＋ si-FSCN1 group（P〈0. 05）. Conclusion Quercetin or FSCN1 gene siRNA can inhibit the vitality of ovarian cancer cells,induce apoptosis and inhibit the expression of VEGF,and the effect of combination is stronger. The mechanism is related to the down regulation of Wnt/β-catenin signaling pathway.