红芪多糖对卵巢癌SKOV3细胞凋亡、生长和运动能力的影响

Effects of radix hedysari polysaccharide on apoptosis,growth and motility of ovarian cancer SKOV3 cells

目的探讨红芪多糖对卵巢癌SKOV3细胞凋亡、生长和运动的调控作用。方法采用0、3、6、12.5、25、50、100、200、400、800、1600 mg/L红芪多糖处理卵巢癌SKOV3细胞,CCK-8法筛选合适剂量,实时荧光定量逆转录聚合酶链反应(qPCR)检测肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)、白介素-6(IL-6)mRNA表达水平;流式细胞术检测细胞凋亡率;克隆形成实验检测细胞生长,Transwell检测细胞侵袭,Western blot检测血管内皮生长因子(VEGF)、上皮型钙黏蛋白(E-cadherin)和纤维粘连蛋白(FN)表达水平。结果选择红芪多糖剂量0、50、100和200 mg/L进行后续实验。与0 mg/L红芪多糖组相比,100 mg/L和200 mg/L红芪多糖组卵巢癌SKOV3细胞中TNF-α、iNOS和IL-6 mRNA表达水平显著下调(P<0.05),细胞凋亡率明显升高(P<0.05),克隆形成率和侵袭细胞数显著降低(P<0.05),E-cadherin蛋白表达上调,VEGF和FN蛋白表达显著下调(P<0.05)。结论红芪多糖能促进卵巢癌SKOV3细胞凋亡,并抑制细胞生长和运动能力。

Objective To investigate the regulatory effect of radix hedysari polysaccharide on the cell apoptosis,growth and motility of ovarian cancer SKOV3 cells.Methods The ovarian cancer SKOV3 cells were treated with 0,3,6,12.5,25,50,100,200,400,800 and 1600 mg/L radix hedysari polysaccharide.CCK8 method was used to screen the appropriate dose.RT-PCR was used to detect tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS)and interleukin-6(IL-6)mRNA expression levels.Flow cytometry was used to detect cell apoptosis.The clone formation test was used to detect cell growth.Transwell method was used to detect cell invasion,and Western blot assay was used to detect the expression levels of vascular endothelial growth factor(VEGF),epithelial cadherin(E-cadherin)and fibronectin(FN).Results The optimal doses of radix hedysari polysaccharide were 0,50,100 and 200 mg/L for the subsequent experiments.Compared with the 0 mg/L radix hedysari polysaccharide group,the expression levels of TNF-α,iNOS and IL-6 mRNA were significantly down-regulated in ovarian cancer SKOV3 cells of 100 mg/L and 200 mg/L radix hedysari polysaccharide groups(P<0.05),the cell apoptotic rate was significantly increased(P<0.05),clone formation rate and the number of invasive cells were significantly decreased(P<0.05),E-cadherin protein expression was up-regulated(P<0.05),and the protein expression levels of VEGF and FN were significantly down-regulated(P<0.05).Conclusion Radix hedysari polysaccharide promotes the apoptosis of ovarian cancer SKOV3 cells and inhibits cell growth and movement.