摘要
目的探讨抗菌肽LL-37对人胃癌细胞系AGS细胞凋亡的影响及其分子机制。方法不同浓度(0、10、20、40μmol/L)LL-37处理胃癌AGS细胞24 h,采用MTT法检测细胞增殖活性,Annexin V-FITC/PI双染流式细胞术法检测细胞凋亡,qRT-PCR法检测细胞中p53 mRNA表达水平,免疫荧光法检测细胞中p53蛋白定位,Western blot法检测细胞核中p53蛋白表达以及细胞中Cleaved-caspase-3、PUMA、Bcl-2和Bax等凋亡相关蛋白表达;干扰p53基因表达验证LL-37通过激活p53信号通路诱导胃癌AGS细胞凋亡。结果LL-37可剂量依赖性抑制胃癌AGS细胞增殖,提高细胞凋亡率,上调p53 mRNA和蛋白表达水平并促进p53蛋白核转位,同时促进凋亡蛋白Cleaved-caspase-3、PUMA和Bax表达,抑制抗凋亡蛋白Bcl-2表达。干扰p53基因表达可抑制LL-37诱导的AGS细胞凋亡。结论LL-37可诱导胃癌AGS细胞凋亡,其机制可能与激活p53表达有关。
Objective To investigate the effect of cathelicidin LL-37 on the apoptosis of gastric cancer AGS cells and its molecular mechanism.Methods Gastric cancer AGS cells were treated with different concentration of LL-37 for 24 h,MTT was used to detect theactivity of cell proliferation,Annexin V-FITC/PI double staining flow cytometry was used to detect cell apoptosis,qRT-PCR method was used to detect the mRNA expression of p53,immunofluorescence method was used to detect the protein localization of p53,Western blot was used to detect the protein expression of p53 in cell nucleus and apoptosis-related proteins such as Cleaned-caspase-3,PUMA,Bcl-2 and Bax in cells;interference with p53 gene expression verified whether LL-37 induced apoptosis of gastric cancer AGS cells by activating p53 signaling.Results LL-37 dose-dependently inhibited the proliferation of gastric cancer AGS cells,elevatedthe rate of apoptosis,up-regulated the mRNA and protein expression level of p53 and promoted the nuclear translocation of p53 protein,while increased the apoptosis proteinsexpression of Cleaved-caspase-3,PUMA and Bax,decreased the anti-apoptotic proteinexpression of Bcl-2.Interference with p53 gene expression inhibited LL-37-induced apoptosis of AGS cells.Conclusion LL-37 can induce apoptosis of gastric cancer AGS cells,and its mechanism may be related to the activation of p53 expression.
作者
欧宏宇
朱海宏
朱文君
贺贝贝
吴世乐
Ou Hongyu;Zhu Haihong;Zhu Wenjun(Dept of General Surgery,Qinghai Provincial People's Hospital,Xining 810007)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第4期571-576,共6页
Acta Universitatis Medicinalis Anhui
基金
青海省卫计委医药卫生科研指导性计划项目(编号:2012-47)。
