岩大戟内酯A和岩大戟内酯B通过Akt信号通路影响MCF-7细胞的增殖和凋亡

摘要

观察岩大戟内酯A(Jolkinolide A,JA)和岩大戟内酯B(Jolkinolide B,JB)二萜类单体对MSC-7乳腺癌细胞的增殖、凋亡影响,并分析JA或JB的作用机制.若分别用40、60、80 μg/mL浓度的JA(或JB)刺激MCF-7细胞,则为不同浓度的JA刺激组或JB刺激组,正常培养的MCF-7为对照组.分别用CCK-8细胞增殖实验检测JA刺激组或JB刺激组的细胞增殖OD值,Western Blot实验检测JA刺激组或JB刺激组MCF-7细胞中Caspase 9蛋白表达,Annexin V-FITC/PI双染检测MCF-7细胞凋亡,酶联免疫吸附实验(ELISA)检测JA刺激组或JB刺激组细胞中Akt、STAT3蛋白的含量.结果发现,与40、60、80 μg/mL的JA刺激组MCF-7细胞分别相比,相应浓度JB刺激组的MCF-7细胞增殖OD值均明显降低,但是JB刺激组MCF-7细胞的Caspase 9蛋白相对吸光度、凋亡率显著升高;相对于40、60、80 μg/mL JA刺激组,相应浓度的JB-刺激组MCF-7细胞上清液中的VEGF含量分别均降低,相应浓度JB刺激组MCF-7细胞的Akt、STAT3蛋白表达均降低.实验结果表明:岩大戟内酯B明显地通过抑制Akt-STAT3信号通路而抑制MCF-7细胞增殖,促进MCF-7凋亡.%To observe the effects of JolkinolideA (JA) and Jolkinolide B (JB) on the proliferation and apoptosis of MCF-7 human breast cancer cells and analyze the mechanism of JA or JB.In this experiment,40,60 and 80 μg/mL of JA or JB were used to stimulate MCF-7 cells,respectively.The cells stimulated by the different concentrations of JA or JB were used as the JA groups or JB groups,respectively.The normal cuhured MCF-7 cells were used as the control group.CCK-8 assay was used to detect the OD value of cell proliferation in JA groups and JB groups;Western blot experiment was used to detect the expression of Caspase 9 protein in JA groups and JB groups;Annexin V-FITC/PI experiment was used to detect the apoptosis rate of MCF-7 cells in JA and JB groups;Enzyme-linked immunosorbent assay (ELISA) experiment was used to detect the contents of Akt and STAT3 protein in JA groups and JB groups.The experimental results found that compared with the JA groups (40,60 and 80 μg/mL),the OD values of MCF-7 cell proliferation in the JB groups with the corresponding concentrations were significantly decreased,and the relative absorbance values of Caspase 9 protein and the cell apoptosis rates in JB groups were significantly increased;Compared with the JA groups,the contents of VEGF in the supernatants of JB groups with the corresponding concentrations were decreased,and the expression of Akt and STAT3 protein were also decreased.These results indicated that JB can promote the apoptosis and inhibit the proliferation of MCF-7 cells by inhibiting the Akt-STAT3 signaling pathway.