芦笋对卵巢癌细胞作用的探究

Effect of Asparagus on Ovarian Cancer Cells

目的利用体外细胞实验初步探究芦笋对卵巢癌细胞的作用及其相关机制。方法选择人高级别浆液性人卵巢癌细胞系MES-OV细胞作为研究对象,比较不同浓度芦笋作用下的人卵巢癌细胞系MES-OV细胞的72 h相对光密度(OD)值;利用荧光光度法测定不同浓度芦笋作用下人卵巢癌细胞系MES-OV细胞线粒体内活性氧释放量;利用酶联免疫吸附法测定不同浓度芦笋作用下人卵巢癌细胞系MES-OV细胞的胱天蛋白酶(caspase)-3的表达量;利用免疫印迹方法测定不同浓度芦笋作用下的人卵巢癌细胞系MES-OV细胞相关凋亡蛋白表达情况。结果不同浓度芦笋作用72 h的MES-OV卵巢癌细胞相对OD值比较差异有统计学意义(P<0.05)。与空白对照组以及0.1 mg/ml芦笋组相比,0.5、1、2.5、5、10 mg/ml芦笋组72 h的MES-OV卵巢癌细胞相对OD值均降低(P<0.05)。与0.5 mg/ml芦笋组比较,1、2.5、5、10 mg/ml芦笋组72 h的MES-OV卵巢癌细胞相对OD值均降低(P<0.05)。与1 mg/ml芦笋组比较,2.5、5、10 mg/ml芦笋组72 h的MES-OV卵巢癌细胞相对OD值均降低(P<0.01)。不同浓度芦笋作用45 min的MES-OV卵巢癌细胞线粒体的活性氧释放量比较差异有统计学意义(P<0.05)。与空白对照组相比,0.1、1、2.5 mg/ml芦笋组MES-OV卵巢癌细胞线粒体的相对活性氧释放量均增加(P<0.05);随着芦笋浓度的升高,MES-OV卵巢癌细胞线粒体的活性氧释放量逐渐增加(P<0.05)。不同浓度芦笋作用16 h的MES-OV卵巢癌细胞培养液中caspase-3相对表达量比较差异有统计学意义(P<0.05)。与空白对照组相比,0.1、1、2.5 mg/ml组MES-OV卵巢癌细胞培养液中caspase-3相对表达量升高(P<0.05);随着芦笋浓度的升高,MES-OV卵巢癌细胞培养液中caspase-3相对表达量(P<0.05)。蛋白印迹结果显示,与空白对照组相比,随着芦笋浓度的升高,MES-OV卵巢癌细胞中Bcl-xL和Mcl-1蛋白表达明显下降。结论芦笋可不同程度抑制卵巢癌细胞增殖并诱导线粒体内活性氧释放,促进细胞凋亡,发挥抗肿瘤作用。

Objective To preliminarily explore the effect of asparagus on ovarian cancer cells and its related mechanism by cell experiment in vitro.Methods Human high-grade serous human ovarian cancer cell line MES-OV was selected as the object of this study,and the 72 h relative optical density(OD)values of the cells under the effect of asparagus at different concentrations were compared;the release of reactive oxygen species in mitochondria of the cells under different concentrations of asparagus was measured by fluorescence spectrophotometry;the expression of caspase-3 in the cells under different concentrations of asparagus was measured by enzyme linked immunosorbent assay method;the expression of apoptosis related proteins in the cells under different concentrations of asparagus was determined by immunoblotting.Results There was significant difference in the relative OD value of MES-OV ovarian cancer cells treated with asparagus at different concentrations for 72 h(P<0.05).Compared with the blank control group and 0.1 mg/ml asparagus group,the relative OD value of the cells in 0.5,1,2.5,5,10 mg/ml asparagus group at 72 h decreased(P<0.05).Compared with 0.5 mg/ml asparagus group,the relative OD value of the cells in 1,2.5,5,10 mg/ml asparagus group decreased at 72 h(P<0.05).Compared with 1 mg/ml asparagus group,the relative OD value of the cells in 2.5,5 and 10 mg/ml asparagus group decreased at 72 h(P<0.01).There was significant difference in the release of reactive oxygen species from mitochondria of the cells treated with asparagus at different concentrations for 45 min(P<0.05).Compared with the blank control group,the release of relative reactive oxygen species in mitochondria of the cells in the 0.1,1 and 2.5 mg/ml asparagus group increased(P<0.05);with the increase of asparagus concentration,the release of reactive oxygen species from mitochondria of the cells gradually increased(P<0.05).There was significant difference in the relative expression of caspase-3 in MES-OV ovarian cancer cell culture medium treated with asparagus at different concentrations for 16 hours(P<0.05).Compared with the blank control group,the relative expression of caspase-3 in the cell culture medium in 0.1,1 and 2.5 mg/ml groups increased(P<0.05);with the increase of asparagus concentration,the relative expression of caspase-3 in the cell culture medium increased(P<0.05).Western blot results showed that compared with the blank control group,the expression of Bcl-xL and Mcl-1 protein in the MES-OV ovarian cancer cells decreased significantly with the increase of asparagus concentration.Conclusion Asparagus can inhibit the proliferation of ovarian cancer cells to varying degrees,induce the release of reactive oxygen species in mitochondria,promote cell apoptosis,and play an anti-tumor role.