萝卜硫素对人结肠癌HT-9细胞增殖、凋亡及PI3K/Akt信号通路的影响

Impacts of sulforaphane on proliferation, apoptosis and PI3K/Akt pathway of human colon cancer HT-9 cells

关键词:  萝卜硫素  人结肠癌细胞  增殖  凋亡  机制

Key words:  sulforaphane  human colon cancer cells  proliferation  apoptosis  mechanism

摘要:

[目的]研究萝卜硫素对人结肠癌HT-9细胞增殖和凋亡的影响及其可能的作用机制。[方法]采用CCK8试剂检测萝卜硫素对HT-9细胞增殖的影响；碘化丙啶（PI）染色检测萝卜硫素对HT-9细胞凋亡的影响；免疫印迹法检测PI3K/Akt信号通路蛋白及凋亡相关蛋白的表达。[结果]萝卜硫素可以明显抑制HT-9细胞的增殖，并呈时间和剂量依赖性；PI染色结果提示萝卜硫素可明显诱导HT-9细胞的凋亡，10、20、40 μg/mL萝卜硫素作用24 h后，HT-9细胞的凋亡率分别为（14.67±1.95）%、（27.95±2.53）%及（35.6±3.75）%，并且让细胞停留在G0/G1期；Western Blot结果提示萝卜硫素呈剂量依赖性的抑制PI3K、p-Akt及Bcl-2蛋白的表达，并诱导Bax蛋白的表达。[结论]萝卜硫素可明显抑制HT-9细胞的增殖并诱导其凋亡，其作用机制可能与抑制PI3K/Akt信号通路的活化有关。

Abstract:

[Objective] To observe the impacts of sulforaphane on proliferation, apoptosis and PI3K/Akt pathway of human colon cancer HT-9 cells.[Methods] The effects of sulforaphane on proliferation of HT-9 cells were investigated using CCK8 assays. The HT-9 cells apoptosis was analyzed by flow cytometry with propidium iodide staining. The expression levels of related proteins in PI3K/Akt pathway and apoptosis proteins were analyzed by Western Blot.[Results] Sulforaphane can significantly suppress proliferation of HT-9 cells in a time-and dose-dependent manner. Propidium iodide staining showed that the apoptosis of HT-9 cells was stimulated by sulforaphane. After treated with 10, 20, 40 μg/mL of sulforaphane for 24 h, the apoptosis rates were (14.67±1.95)%, (27.95±2.53)% and (35.6±3.75)%, respectively. Sulforaphane downregulated the expression levels of PI3K, p-Akt and Bax proteins and upregulated the expression of Bax protein in a dose-dependent manner.[Conclusion] Sulforaphane can significantly suppress the proliferation of HT-9 cells and stimulate apoptosis, the mechanism may be associated with the inhibition of PI3K/Akt signaling pathway.