摘要
目的探讨金合欢素(Aca)通过调节PTEN诱导激酶1(PINK1)/E3泛素连接酶(Parkin)通路介导的线粒体自噬对肝癌HepG2细胞增殖、凋亡和迁移的影响。方法将肝癌HepG2细胞分为对照组(正常培养的HepG2细胞)、Aca组(10μmol/L Aca)、PINK1小干扰RNA阴性对照(si-NC)组(转染si-NC)、PINK1小干扰RNA(si-PINK1)组(转染si-PINK1)、Aca+si-NC组(转染si-NC后用10μmol/L Aca处理)、Aca+si-PINK1组(转染si-PINK1后用10μmol/L Aca处理)。CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡;Transwell实验检测细胞迁移;透射电镜观察自噬小体的形成;Western blot检测细胞中自噬相关蛋白[Beclin-1、微管相关蛋白1轻链3(LC3)-Ⅰ、LC3-Ⅱ]及PINK1/Parkin通路相关蛋白表达。结果与对照组比较,Aca组HepG2细胞存活率、迁移细胞数目降低,凋亡率、自噬小体数量、Beclin-1、LC3-Ⅱ/LC3-Ⅰ、PINK1、Parkin蛋白表达水平升高(P<0.05),si-PINK1组HepG2细胞存活率、迁移细胞数目升高,凋亡率、自噬小体数量、Beclin-1、LC3-Ⅱ/LC3-Ⅰ、PINK1、Parkin蛋白表达水平降低(P<0.05);与Aca组、Aca+si-NC组比较,Aca+si-PINK1组HepG2细胞存活率、迁移细胞数目升高,凋亡率、自噬小体数量、Beclin-1、LC3-Ⅱ/LC3-Ⅰ、PINK1、Parkin蛋白表达水平降低(P<0.05)。结论Aca可能通过激活PINK1/Parkin通路介导的线粒体自噬抑制肝癌HepG2细胞增殖、迁移,促进细胞凋亡。
Objective To investigate the effect of acacetin(Aca)on the proliferation,apoptosis and migration of liver cancer HepG2 cells by regulating PTEN induced kinase 1(PINK1)/E3 ubiquitin protein ligase(Parkin)pathway mediated mitochondrial autophagy.Methods Liver cancer HepG2 cells were divided into the control group(normally cultured HepG2 cells),the Aca group(10μmol/L Aca),the PINK1 small interfering RNA negative control(si-NC)group(transfected with si-NC),the PINK1 small interfering RNA(si-PINK1)group(transfected with si-PINK1),the Aca+si-NC group(treated with 10μmol/L Aca after transfection of si-NC)and Aca+si-PINK1 group(treated with 10μmol/L Aca after transfection of si-PINK1).Cell counting kit-8(CCK-8)method was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Transwell test was used to detect cell migration.Transmission electron microscope was used to observe the formation of autophagosomes.Western blot assay was used to detect expression levels of autophagy related proteins[Beclin-1,microtubule associated protein 1 light chain 3(LC3)-Ⅰand LC3-Ⅱ]and PINK1/Parkin pathway related proteins in cells.Results Compared with the control group,the survival rate and the number of migrating of HepG2 cells were significantly decreased in the Aca group,and the apoptosis rate,number of autophagy bodies,expression levels of Beclin-1,LC3-Ⅱ/LC3-Ⅰ,PINK1 and Parkin protein were increased significantly(P<0.05).The survival rate and the number of migrating of HepG2 cells were significantly increased in the si-PINK1 group,and the apoptosis rate,number of autophagy bodies,expression levels of Beclin-1,LC3-Ⅱ/LC3-Ⅰ,PIN1 and Parkin protein were decreased significantly(P<0.05).Compared with the Aca group and the Aca+si-NC group,the survival rate and the number of migrating of HepG2 cells were significantly increased in the Aca+si-PINK1 group,and the apoptosis rate,number of autophagy bodies,expression levels of Beclin-1,LC3-Ⅱ/LC3-Ⅰ,PINK1 and Parkin protein were decreased significantly(P<0.05).Conclusion Aca may inhibit the proliferation and migration and promote cell apoptosis of liver cancer HepG2 cells by activating mitochondrial autophagy mediated by PINK1/Parkin pathway.
作者
吴琼
李锦源
黄文涛
安娜
WU Qiong;LI Jinyuan;HUANG Wentao;AN Na(Department of Pharmacy,Cancer Hospital Affiliated to Guangxi Medical University,Nanning 530021,China)
出处
《天津医药》
CAS
北大核心
2023年第3期235-239,共5页
Tianjin Medical Journal
基金
广西壮族自治区中青年教师基础能力提升项目(KY2016LX037)。
