芒柄花黄素对人鼻咽癌细胞株HONE1凋亡的诱导作用及其机制探讨

摘要

目的 观察芒柄花黄素对人鼻咽癌细胞株HONE1细胞凋亡的诱导作用,并探讨其相关机制.方法 用含不同浓度(0、5、10、20、40、80、160μmol/L)芒柄花黄素的培养基分别培养人鼻咽癌细胞株HONE124、48、72 h,采用MTT法观察芒柄花黄素对HONE1细胞活性的影响.采用Hoechst33258荧光染色从形态学上观察芒柄花黄素处理的HONE1细胞凋亡情况.用Western blotting检测不同浓度(0、5、10、20、40μmol/L)芒柄花黄素处理的HONE1细胞的p-Akt、Akt、BAX和Bcl-2蛋白.结果 Hoechst33258荧光染色结果显示,芒柄花黄素处理较未处理的人鼻咽癌细胞株HONE1凋亡细胞数明显增加.0、5、10、20、40μmol/L芒柄花黄素处理48 h后HONE1细胞p-Akt/Akt值分别为0.311±0.035、0.169±0.029、0.116±0.038、0.143±0.012、0.082±0.015,5、10、20、40μmol/L与0μmol/L比较,P均＜0.05.0、5、10、20、40μmol/L芒柄花黄素处理48 h后HONE1细胞BAX/Bcl-2值分别为1.445±0.167、1.678±0.257、2.637±0.619、3.292±0.339、4.285±0.899,10、20、40μmol/L与0μmol/L比较,P均＜0.05.结论 芒柄花黄素可诱导人鼻咽癌细胞株HONE1细胞凋亡,这可能是通过下调p-Akt/Akt、上调BAX/Bcl-2实现的.%Objective To observe the induction of formononetin on apoptosis of human nasopharyngeal carcinoma cell line HONE1 and to investigate its mechanism .Methods HONE1 cells were cultured with different concentrations of form-ononetin (0, 5, 10, 20, 40, 80 and 160 μmol/L) for separated time (24, 48 and 72 h).Cell viability of HONE1 was examined using an MTT assay .The apoptosis of HONE1 cells from morphological changes after being treated with form-ononetin was examined by Hoechst33258 assay.The proteins (p-Akt, Akt, BAX and Bcl-2) of HONE1 cells were tested by Western blotting after being treated with different concentrations of formononetin (0, 5, 10, 20 and 40μmol/L) for 48 h. Results Hoechst33258 fluorescence staining showed that the number of apoptosis of HONE 1 cells was increased after being treated with formononetin as compared with the cells without treatment .The p-Akt/Akt values of HONE1 cells which were treated with different concentrations of formononetin (0, 5, 10, 20 and 40 μmol/L) for 48 h were 0.311 ±0.035, 0.169 ± 0.029, 0.116 ±0.038, 0.143 ±0.012 and 0.082 ±0.015, respectively.Significant difference was found between 0μmol/L and 5, 10, 20 and 40μmol/L (all P＜0.05).The BAX/Bcl-2 values of HONE1 cells which were treated with different con-centrations of formononetin (0, 5, 10, 20 and 40μmol/L) for 48 h were 1.445 ±0.167, 1.678 ±0.257, 2.637 ±0.619, 3.292 ±0.339 and 4.285 ±0.899, respectively.Significant difference was found between 0 μmol/L and 10, 20 and 40μmol/L (all P＜0.05).Conclusion Formononetin can induce the apoptosis of human nasopharyngeal carcinoma cell line HONE1, which may be mediated by down-regulating p-Akt/Akt and up-regulating BAX/Bcl-2.