摘要
目的探讨黄腐醇对人胃癌细胞系SGC-7901焦亡的作用及其可能机制。方法用DMSO(对照组)和不同浓度黄腐醇10、20和40μmol/L处理SGC-7901细胞,采用细胞集落实验检测细胞集落形成,细胞划痕实验观察细胞迁移,流式细胞术检测细胞内活性氧(ROS)水平,Western blot法检测加斯德明E-N端(GSDME-N)和Caspase-3蛋白表达。采用ROS清除剂N-乙酰-L-半胱氨酸(NAC)5 mmol/L、细胞外信号调节激酶(ERK)抑制剂U012610μmol/L或p38抑制剂达马莫德(BIRB)10μmol/L预处理SGC-7901细胞后再加入黄腐醇40μmol/L,采用Western blot法检测GSDME-N蛋白表达,倒置显微镜下观察细胞焦亡的形态学变化。结果与对照组相比,黄腐醇呈浓度依赖性地抑制SGC-7901细胞集落形成、细胞迁移和Caspase-3蛋白表达,上调ROS水平和GSDME-N蛋白表达(P<0.05);而用NAC、U0126或BIRB预处理后,GSDME-N蛋白表达降低,细胞焦亡数目减少(P<0.05)。结论黄腐醇可能通过升高ROS水平和作用ERK/p38通路诱导SGC-7901细胞焦亡。
Objective To investigate the effect and underlying mechanism of xanthohumol on the pyroptosis of gastric cancer cell line SGC-7901.Methods SGC-7901 cells were treated with DMSO(control group)and different concentrations of xanthohumol 10,20 and 40μmol/L.The formation of cell colonies was determined by cell colony experiment,and the cell migration was measured by wound healing assay.The level of reactive oxygen species(ROS)was analyzed by flow cytometry,and the protein expressions of Gasdermin E-N terminal(GSDME-N)and Caspase-3 were detected by Western blot.SGC-7901 cells were preincubated with ROS scavenger N-acetyl-L-cysteine(NAC)5 mmol/L,extracellular signal-regulated kinase(ERK)inhibitor U012610μmol/L or p38 inhibitor Doramapimod(BIRB)10μmol/L and then treated with xanthohumol 40μmol/L.The protein expression of GSDME-N was determined by Western blot,and the morphological change of cell pyroptosis was observed under inverted microscope.Results Compared with control group,xanthohumol inhibited the formation of cell colony,cell migration and protein expression of Caspase-3,and upregulated the ROS level and protein expression of GSDME-N in a concentration-dependent manner(P<0.05).While the protein expression of GSDME-N and the number of cell pyroptosis were decreased after the preincubation of NAC,U0126 or BIRB(P<0.05).Conclusion Xanthohumol could induce the pyroptosis of SGC-7901 cells by increasing ROS level and acting on the ERK/p38 pathway.
作者
叶甜甜
李震岳
吴守彪
周陈建
翁约约
苏银法
YE Tiantian;LI Zhenyue;WU Shoubiao(Department of Pharmacy,Wenzhou Central Hospital,Wenzhou 325000,CHINA)
出处
《江苏医药》
CAS
2022年第3期222-226,230,共6页
Jiangsu Medical Journal
基金
温州市基础性科研项目(Y2020181)。
关键词
胃癌
黄腐醇
细胞焦亡
活性氧
细胞外信号调节激酶/p38通路
Gastric cancer
Xanthohumol
Cell pyroptosis
Reactive oxygen species
Extracellular signal-regulated kinase/p38 pathway