Objective To observe the effects of formononetin on the proliferation and apoptosis of human gastric cancer MKN-45 cells and its possible mechanism.Methods The human gastric cancer MKN-45 cells in the logarithmic growth phase were divided into groups A,B,C and the control group.Cells in the groups A,B and C were cultured and the culture medium was added with 20,40 and 80 μg/mL of formononetin,respectively.The control group was not treated.The cell proliferation of MKN-45 cells was detected by MTT assay at 24,48 and 72 h of formononetin treatment,and the growth inhibitory rate of MKN-45 cells was assessed.The effect of formononetin on morphological changes was evaluated using DAPI staining.The intracellular phosphorylated IκB (p-IκB) and NF-κB p65 protein expression was measured by Western blotting.Results MKN-45 cell proliferation was inhibited after being treated by formononetin with a time-and dose-dependent manner (all P ＜ 0.05).The apoptosis rates of groups A,B,C and the control group after 72-hour treatment were 25.62％ ± 2.57％,48.27％ ± 3.18 ％,72.51％ ± 4.35％ and 1.07％ ± 0.54％,respectively (all P ＜ 0.05).The expression of p-IκB and NF-κB p65 protein in cells of groups A,B and C was significantly higher than that of the control group at 48 h after administration with a dose-dependent manner (all P ＜ 0.05).Conclusion Formononetin inhibits cell proliferation and induces apoptosis of MKN-45 cells,and the possible mechanism may be related to activation of NF-κB signaling pathway.%目的 观察芒柄花黄素对人胃癌MKN-45细胞株增殖、凋亡的影响,并探讨其可能作用机制.方法 取对数生长期人胃癌MKN-45细胞分为A、B、C、对照组,A、B、C组分别加入20、40、80 μg/mL芒柄花黄素培养,对照组加入不含芒柄花黄素的完全培养基.采用MTT法观察给药24、48、72 h各组细胞增殖情况,计算细胞增殖抑制率.采用DAPI染色法评价其对MKN-45细胞形态学的影响.采用Western blotting法检测给约48 h时各组细胞磷酸化IκB(p-IκB)及NF-κB p65.结果 随芒柄花黄素浓度升高,细胞增殖抑制率升高,且随干预时间增加,细胞增殖抑制率升高(P均＜0.05).给药72 h时A、B、C、对照组的凋亡率分别为25.62％±2.57％、48.27％±3.18％、72.51％±4.35％、1.07％±0.54％,A、B、C组与对照组相比,P均＜0.05.给药48 h时A、B、C组细胞p-IκB、NF-κB p65蛋白相对表达量均高于对照组,且随芒柄花黄素浓度升高,细胞p-IκB及NF-κB p65蛋白相对表达量增加,P均＜0.05.结论 芒柄花黄素可抑制人胃癌MKN-45细胞株的增殖、促进细胞凋亡,其机制可能为芒柄花黄素激活NF-κB信号通路.
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