萝卜硫素介导Wnt/β-catenin信号通路诱导胃癌细胞凋亡的研究

Study on the apoptosis of gastric cancer cells induced by sulforaphane mediated Wnt/β-catenin signaling pathway

关键词:  萝卜硫素  胃癌细胞  增殖  凋亡  Wnt/β-catenin信号通路

Key words:  sulforaphane  gastric cancer cell  proliferation  apoptosis  Wnt/β-cateni signaling pathway

摘要:

[目的]探讨萝卜硫素对胃癌SGC-7901细胞增殖的影响及其对Wnt/β-连锁蛋白（Wnt/β-catenin）信号通路活化的影响。[方法]采用CCK8实验检测10、20、40 μmol/L萝卜硫素对SGC-7901细胞增殖的影响，流式细胞仪分析萝卜硫素对SGC-7901细胞凋亡的影响，荧光素酶报告基因实验检测萝卜硫素对Wnt/β-catenin通路中淋巴增强因子/淋巴腺黏连因子（TCF4/LEF）转录活性的影响，Western Blot方法检测萝卜硫素对SGC-7901细胞中β-catenin、糖原合成激酶3β（GSK-3β）、c-Myc及半光天冬酶3（Caspase3）蛋白表达水平的影响。[结果]与对照组相比，萝卜硫素组SGC-7901细胞增殖抑制率显著升高（P<0.05），呈现出时间、剂量依赖性；萝卜硫素组SGC-7901细胞凋亡率相比对照组显著增高（P<0.05），并且萝卜硫素还能明显抑制TCF4/LEF报告质粒的荧光素酶活性（P<0.05）；萝卜硫素组SGC-7901细胞内β-catenin及c-Myc蛋白表达呈浓度依赖性降低，而GSK-3β及Caspase3蛋白表达呈浓度依赖性增高（P<0.05）。[结论]萝卜硫素能明显抑制SGC-7901细胞增殖，并诱导其凋亡，其作用机制可能与抑制Wnt/β-catenin信号通路活化有关。

Abstract:

[Objective] To investigate the effect of sulforaphane on the proliferation of gastric cancer SGC-7901 cells and its effect on the activation of Wnt/β-catenin signaling pathway.[Methods] The effects of 10,20,40 μmol/L sulforaphane on the proliferation of SGC-7901 cells were detected by CCK8 assay. The effect of sulforaphane on the apoptosis of SGC-7901 cells was analyzed by flow cytometry. The luciferase reporter gene assay was used to detect the effect of sulforaphane on the transcriptional activity of lymphatic enhancement factor/lymphatic adhesion factor (TCF4/LEF) in Wnt/β-catenin pathway. Western Blot method was used to detect the effect of sulforaphane on the β-catenin,glycogen synthase kinase 3β,c-Myc and Caspase 3 protein expression levels in SGC-7901 cells.[Results] Compared with the control group,the inhibition rate of SGC-7901 cells in the sulforaphane group was significantly increased (P<0.05),showing a time-and dose-dependent manner;the apoptosis rate of SGC-7901 cells in the sulforaphane group was significantly higher than that in the control group (P<0.05),and sulforaphane could also significantly inhibit the luciferase activity of TCF4/LEF reporter plasmid (P<0.05). The expression of β-catenin and c-Myc protein in SGC-7901 cells of sulforaphane group was concentration-dependent increase,while the expression of GSK-3β and Caspase3 protein increased in a concentration-dependent manner (P<0.05).[Conclusion] The sulforaphane can significantly inhibit the proliferation and induce apoptosis of SGC-7901 cells,and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway activation.