萝卜硫素通过Traf6/TAK1信号通路介导人胃癌细胞凋亡的机制研究

Mechanism study of sulforaphane on cell apoptosis in human gastric cancer cells through Traf6/TAK1 signaling pathway

关键词:  萝卜硫素  人胃癌细胞  增殖  凋亡  机制

Key words:  sulforaphane  human gastric cancer cell  proliferation  apoptosis  mechanism

摘要:

[目的]探讨萝卜硫素对人胃癌（HGC27）细胞增殖、凋亡的影响，并观察其可能作用的机制。[方法]将对数生长期的HGC27细胞分为空白对照组、15 μg/mL萝卜硫素、30 μg/mL萝卜硫素及60 μg/mL萝卜硫素，用不同浓度药物处理细胞后，采用CCK8法检测各组细胞的增殖情况，流式细胞术检测各组细胞的凋亡率，免疫印迹技术检测细胞中凋亡蛋白及肿瘤坏死因子受体相关分子6（Traf6）/转化生长因子-β活化激酶1（TAK1）信号通路蛋白表达量。[结果]与空白对照组相比，不同浓度萝卜硫素组细胞增殖抑制率及凋亡率明显升高（P<0.05）；促凋亡蛋白半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）、Bcl-2相关X蛋白（Bax）表达水平显著升高，而抗凋亡蛋白B淋巴细胞瘤-2（Bcl-2）水平则显著降低（P<0.05）；Traf6、p-TAK1蛋白表达量相对于空白对照组显著升高（P<0.05）。[结论]萝卜硫素可以抑制人胃癌细胞的增殖，促进细胞的凋亡，Traf6/TAK1信号通路可能参与了萝卜硫素对肿瘤细胞的抑制作用。

Abstract:

[Objective] To study the impacts of sulforaphane on the apoptosis of human gastric cancer HGC27 cells and its roles in Traf6/TAK1 signaling pathway, and to verify the mechanisms.[Methods] The HGC27 cells were divided into control group, low dose of sulforaphane group (15 μg/mL), middle dose of sulforaphane group (30 μg/mL), and high dose of sulforaphane group (60 μg/mL). After cells were supplemented with varying doses of sulforaphane, the inhibitory rates of cell proliferation were determined using CCK8 assay, the apoptotic rates were assessed using flow cytometry, and the expression levels of Caspase-3, Bax, Bcl-2, Traf6, TAK1 and p-TAK1 were observed by Western Blot.[Results] Compared with control group, the inhibitory rates of proliferation and the apoptotic rates of cells in different sulforaphane doses groups were greatly increased (P<0.05); the expression levels of Caspase-3 and Bax were increased while the level of Bcl-2 was greatly decreased compared with control group (P<0.05); the expression levels of Traf6, p-TAK1 were also greatly increased (P<0.05).[Conclusions] Sulforaphane can suppress the proliferation of HGC27 cells and induce the apoptosis. Traff6/TAK1 signaling pathway may be involved in the inhibitory effect of sulforaphane on the tumor cells.