萝卜硫素对视网膜母细胞瘤细胞Wnt/β-catenin信号通路的影响

Impacts of sulforaphane on the Wnt/β-catenin pathway of cultured retinoblastoma cells

目的 研究萝卜硫素对视网膜母细胞瘤细胞(Y79)增殖、凋亡及Wnt/β-catenin信号通路的影响。方法Y79细胞经10、20、40 mg/L萝卜硫素干预24、48 h后,采用CCK8试剂盒检测细胞增殖抑制率,采用流式细胞仪检测萝卜硫素对Y79细胞凋亡的影响,Western blot法检测凋亡相关蛋白Bax、Bcl-2及Wnt/β-catenin信号通路相关蛋白的表达水平。结果不同浓度萝卜硫素作用Y79细胞24、48 h后,可呈浓度、时间依赖性的抑制细胞增殖。与对照组比较,10、20、40 mg/L萝卜硫素组细胞凋亡率[(12.47±2.24)%、(24.63±3.44)%、(57.49±5.41)%比(3.47±0.74)%]增高(P<0.05);40 mg/L萝卜硫素组细胞Bax[(0.57±0.03)比(0.12±0.01)]表达升高,抗凋亡蛋白Bcl-2[(0.16±0.01)比(0.81±0.03)]、Wnt[(0.19±0.01)比(0.74±0.02)]、β-catenin[(0.22±0.02)比(0.82±0.03)]蛋白表达降低(P<0.05)。结论萝卜硫素可抑制Y79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。

Objective To observe the impacts of sulforaphane on proliferation, apoptosis and Wnt/β-catenin pathway of cultured retinoblastoma cells. Methods Retinoblastoma Y79 cells were cultured with varying concentrations of sulforaphane (10, 20, 40 mg/L) for 24 and 48 h. Cell proliferation and apoptosis were measured by CCK8 assay and flow cytometry. Apoptosis related proteins Bax and Bcl-2, as well as the expression levels of related proteins in Wnt/β-catenin signaling pathway were determined using Western Blot. Results Sulforaphane suppressed cell proliferation and stimulate apoptosis (12.47% ± 2.24%, 24.63% ± 3.44%, 57.49% ± 5.41% vs. 3.47% ± 0.74%) in a dose- and time-dependent manner, respectively. Furthermore, 40 mg/L of sulforaphane increased the expression of Bax (0.57 ± 0.03 vs. 0.12 ± 0.01), but decreased the Bcl-2 (0.16 ± 0.01 vs. 0.81 ± 0.03) protein expression. The results of Western Blot displayed that sulforaphane decreased the expression levels of Wnt (0.19 ± 0.01 vs. 0.74 ± 0.02), β-catenin protein (0.22 ± 0.02 vs. 0.82 ± 0.03), indicating that sulforaphane can inhibit the activation of Wnt/β-catenin signaling pathway. Conclusions The sulforaphane could inhibit the proliferation and induce the apoptosis of retinoblastoma Y79 cells, probably regulating by the Wnt/β-catenin signaling pathway.