布雷菲德菌素A对HepG2细胞自噬的诱导作用  

Study of brefeldin a on induction of autophagy in HepG2 Cells

作　　者：李丹 李明虹 李星道 刘希冲 高文婷 余红梅 白剑英 LI Dan;LI Minghong;LI Xingdao;LIU Xichong;GAO Wenting;YU Hongmei;BAI Jianying(Department of Environmental Health,School of Public Health,Shanxi Medical University,Taiyuan,030001,China;Department of Health Statistics,School of Public Health,Shanxi Medical University,Taiyuan,030001,China)

机构地区：[1]山西医科大学公共卫生学院环境卫生学教研室,太原030001 [2]山西医科大学公共卫生学院卫生统计学教研室,太原030001

出　　处：《环境化学》2022年第10期3345-3352,共8页Environmental Chemistry

摘　　要：为探讨新型抗癌药物布雷菲德菌素A(BFA)诱导肝癌细胞发生自噬性死亡的作用机制,以HepG2细胞作为研究对象,将细胞分成阴性对照组、不同浓度BFA组和阳性对照组(雷帕霉素),采用MTT法测定HepG2细胞活性;采用免疫荧光法检测细胞内自噬标志蛋白LC3的表达和细胞核形态学改变;采用Western Blot法检测细胞内激活转录因子4(ATF4)、C/EBP同源蛋白(CHOP)、聚(腺苷二磷酸-核糖)多聚酶(PARP)、增殖细胞核抗原(PCNA)、微管相关蛋白1轻链3(LC3)、自噬相关蛋白5(Atg5)、自噬相关蛋白6(Atg6/Beclin-1)的表达水平.结果表明,BFA可引起HepG2细胞内自噬特征蛋白LC3-Ⅱ、Atg5、Beclin-1的蛋白表达水平明显增加;同时引起HepG2细胞活性降低、细胞数量减少、细胞核固缩和变形.说明BFA可能引起HepG2细胞发生自噬性细胞死亡.进一步探讨自噬诱导途径发现,BFA引起HepG2细胞内ATF4、CHOP的蛋白表达水平增加.BFA可能通过诱导ATF4和CHOP的过表达引起HepG2细胞发生自噬性细胞死亡.To investigate the mechanism of the new anticancer drug Brefeldin A(BFA)induced autophagic cell death in HepG2 cells.MTT assay was used to measure the activity of HepG2 cells.The immunofluorescence method was used to observed the expression of LC3 and nuclear morphological changes.Measurement of activating transcription factor 4(ATF4),C/EBP homologous protein(CHOP),poly ADP-ribose polymerase(PARP),proliferating cell nuclear antigen(PCNA),microtubule-associated protein 1 light chain 3(LC3),autophagy related 5 homolog(Atg5),autophagy related 6 homolog(Atg6/Beclin-1)by Western blot.The results showed that BFA could significantly increase the expression levels of autophagy characteristic proteins LC3-Ⅱ,Atg5 and Beclin-1 in HepG2 cells;At the same time,it caused the decrease of HepG2 cell activity,cell number reduction,nucleus pyknosis and deformation.These results suggest that BFA may cause autophagic cell death in HepG2 cells.Further exploration of the autophagy induction pathway found that BFA caused an increase in the protein expression levels of ATF4 and CHOP in HepG2 cells.The above results suggest that BFA may induce autophagic cell death in HepG2 cells by inducing the overexpression of ATF4 and CHOP.

关 键 词：布雷菲德菌素A HEPG2细胞 激活转录因子4 C/EBP同源蛋白 自噬性细胞死亡 

分 类 号：R965[医药卫生—药理学]