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目的探讨表没食子儿茶素没食子酸酯(EGCG)对乳腺癌细胞MCF-7增殖和凋亡的影响,并研究其可能的Notch1-Hes1信号通路机制。方法分别以不同浓度EGCG(20μg/ml、80μg/ml、120μg/ml、160μg/ml)干预MCF-7细胞,采用CCK-8法检测EGCG对MCF-7细胞增殖的抑制作用,流式细胞术法检测EGCG对MCF-7细胞凋亡的影响,并分别采用实时荧光定量PCR和Western blot法检测EGCG干预后MCF-7细胞中Notch1mRNA及Notch1、Hes1蛋白表达的变化。结果 EGCG抑制MCF-7细胞增殖的最佳有效浓度为160μg/ml,作用24、48 h后,观察组细胞增殖抑制率高于空白组(P<0.01),观察组细胞增殖抑制率与阳性对照组比较,差异无统计学意义(P>0.05)。经不同浓度EGCG干预MCF-7细胞48 h后,随着EGCG剂量的增加,MCF-7细胞凋亡率增高。EGCG可不同程度地下调MCF-7细胞中Notch1 mRNA的表达,呈现剂量依赖性趋势,EGCG可影响MCF-7细胞中Notch1、Hes1蛋白表达,与空白组比较,20μg/ml、80μg/ml EGCG可增加Notch1蛋白表达,120μg/ml、160μg/ml EGCG可降低Notch1蛋白表达,其中160μg/ml EGCG可降低Notch1蛋白表达(P<0.05);20μg/ml EGCG增加Hes1蛋白表达量,80μg/ml、120μg/ml、160μg/ml EGCG可降低Hes1蛋白表达量,其中160μg/ml EGCG能够明显降低Hes1蛋白表达(P<0.01)。结论 EGCG可有效抑制乳腺癌细胞MCF-7的增殖,并诱导其凋亡,其作用机制可能与EGCG影响Notch1-Hes1信号通路有关。
Objective To investigate the effect of epigallocatechin-3-gallate (EGCG) on the proliferation and apoptosis of breast cancer cell line MCF-7 and investigate the possible mechanism of Notch1-Hes1 signaling pathway. Methods MCF-7 cells were treated with different concentrations of EGCG (20μg / ml, 80μg / ml, 120μg / ml and 160μg / ml) respectively. The inhibitory effect of EGCG on the proliferation of MCF-7 cells was detected by CCK- The changes of Notch1 mRNA and Notch1, Hes1 protein in MCF-7 cells were detected by real-time fluorescence quantitative PCR and Western blot respectively. Results The best effective concentration of EGCG to inhibit the proliferation of MCF-7 cells was 160μg / ml. After 24 and 48 hours, the inhibition rate of EGCG in the observation group was higher than that in the blank group (P <0.01) There was no significant difference between the two groups (P> 0.05). The apoptosis rate of MCF-7 cells increased with the increase of EGCG dose after 48 h intervention of EGCG at different concentrations. EGCG could down-regulate the expression of Notch1 mRNA in MCF-7 cells to a certain extent in a dose-dependent manner. EGCG could affect the expression of Notch1 and Hes1 in MCF-7 cells. Compared with the blank control group, EGCG at 20μg / ml and 80μg / ml EGCG Increasing Notch1 protein expression at 120μg / ml and 160μg / ml EGCG decreased Notch1 protein expression, while 160μg / ml EGCG decreased Notch1 protein expression (P <0.05); 20μg / ml EGCG increased Hes1 protein expression at 80μg / ml and 120μg / ml and 160μg / ml EGCG could reduce the expression of Hes1 protein, and 160μg / ml EGCG could significantly reduce the expression of Hes1 protein (P <0.01). Conclusion EGCG can effectively inhibit the proliferation and induce the apoptosis of MCF-7 breast cancer cells, which may be related to the effect of EGCG on Notch1-Hes1 signaling pathway.