AMP依赖蛋白激酶叉头转录因子O3a信号通路及氧化应激介导紫铆因诱导人食管癌细胞凋亡作用研究.docVIP

AMP依赖蛋白激酶叉头转录因子O3a信号通路及氧化应激介导紫铆因诱导人食管癌细胞凋亡作用研究 　　[摘要] 目的 探讨紫铆因（Butein）诱导人食管癌EC109细胞凋亡的作用及AMP依赖的蛋白激酶（AMPK）/叉头转录因子O3a（FOXO3a）信号通路、氧化应激在该过程中发挥的作用。 方法 常规培养EC109细胞，分别给予Butein处理，首先检测EC109细胞的活力、迁移能力、氧化应激水平及凋亡指标Caspase 3的活性。采用Western blot法检测细胞内AMPK、FOXO3a的磷酸化水平及凋亡调节蛋白Bim、Bcl2相关X蛋白（Bax）的表达水平。用Compound C阻断AMPK后检测相关指标。建立裸鼠皮下肿瘤模型，给予Butein和Compound C处理，检测裸鼠体重和肿瘤体积。 结果 Butein处理后，细胞活力下降（P 0.05）；细胞间距增加（P 0.05）；Caspase 3活性、NADPH氧化酶活性、ROS含量升高（P 0.05）；总GSH下降，提示凋亡增加、氧化应激增强；p-AMPK磷酸化水平、p-FOXO3a磷酸化水平升高（P 0.05）；Bim、Bax表达升高（P 0.05）；Compound C处理逆转了这一过程。裸鼠皮下肿瘤检测发现Butein抑制在体肿瘤增殖，阻断AMPK可以逆转该抑制作用。 结论 Butein能显著促进食管癌EC109细胞凋亡。该作用可能是通过激活AMPK/FOXO3a信号通路来实现的。 　　[关键词] 紫铆因；食管癌；AMP依赖的蛋白激酶；叉头蛋白转录因子O3a；氧化应激；凋亡 　　[中图分类号] R735.1 [文献标识码] A [文章编号] 1673-7210（2017）06（c）-0025-05 　　[Abstract] Objective To investigate the effects of Butein on esophagus cancer EC109 cells and the role of AMP-activated protein kinase （AMPK）/forkhead class box O3a （FOXO3a） and oxidative stress in this process. Methods EC109 cell was routine cultured and given Butein， the cell vitality， migration ability， cellular oxidative stress level of EC109 cell and Caspase 3 activity were detected. The AMPK， FOXO3 aphosphorylation level and the expressions of Bim and Bax were detected by Western blot. After Compound C inhibit AMPK， related indicators were detected. The nude mice tumor bearing model was made， Butein and Compound C were given， the weight and tumor volume of nude mice were detected. Results After Butein treatment， cell vitality reduced， the distance between cell boundaries increased， ROS concentration and NADPH oxidase activity increased， total GSH level and Caspase 3 activity reduced （P 0.05）； AMPKand FOXO3 aphosphorylation and cell apoptosis increased （P 0.05）； Compound C treatment reversed this process. The nude mice tumor study showed that Butein inhibited the tumor growth and Compound C reversed this effect. Conclusion Butein can induce EC109 cell apoptosis， and this process may be mediated by activation of AMPK/F