ATP5J通过TOMM20调节线粒体功能并促进人肝细胞癌细胞转移  

ATP5J regulates mitochondrial function through TOMM20 and promotes metastasis of human hepatocellular carcinoma cells

作　　者：冷君志 王根旺[1] 刘迪[1] 柳科军 王琦[1] 惠永峰[1] LENG Junzhi;WANG Genwang;LIU Di;LIU Kejun;WANG Qi;HUI Yongfeng(Department of Hepatobiliary Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院肝胆外科,宁夏银川750004

出　　处：《中国病理生理杂志》2024年第3期431-437,共7页Chinese Journal of Pathophysiology

基　　金：宁夏回族自治区重点研发计划项目(No.2021BEG03067);宁夏自然科学基金资助项目(No.2022AAC03545)。

摘　　要：目的:探讨ATP合成酶H+转运线粒体F0复合体亚基F6(ATP5J)通过调节肝癌细胞线粒体功能介导细胞骨架重塑影响肝癌细胞转移的作用及其机制。方法:培养人肝细胞癌Li-7细胞株,基因修饰ATP5J表达(过表达和敲减)。使用JC-1染色检测每组细胞线粒体膜电位情况;利用DCFH-DA荧光探针检测Li-7细胞的活性氧(ROS)含量;线粒体ATP荧光探针检测线粒体功能;微丝绿色荧光探针(Actin-Tracker Green-488)检测细胞骨架重塑情况;Transwell检测细胞侵袭能力;Western blot检测ATP5J和线粒体外膜转位酶20(TOMM20)的表达水平。结果:过表达ATP5J可上调线粒体膜电位水平和线粒体ATP荧光强度、诱导细胞骨架重塑、促进细胞侵袭和TOMM20蛋白表达,抑制ROS生成(P<0.01)。相反,敲减ATP5J显著降低线粒体膜电位和线粒体ATP荧光强度,显著降低细胞侵袭能力和TOMM20表达,促进ROS产生,阻断细胞骨架重塑(P<0.01)。结论:ATP5J可调节肝癌细胞线粒体能量转化,其通过TOMM20调节线粒体膜电位水平和线粒体ATP产生介导细胞骨架重塑影响肝癌细胞转移。AIM:To explore the mechanism of ATP synthase mitochondrial F0 complex H+transporting,subunit F6(ATP5J)in affecting the metastasis of hepatoma carcinoma cells by regulating mitochondrial function-mediated cytoskeletal remodeling.METHODS:Hepatocellular carcinoma cells Li-7 were used to construct the ATP5J overexpression and knockdown models.JC-1 staining was used to detect the mitochondrial membrane potential in each group,reactive oxygen species(ROS)levels were examined by DCHF-DA,and mitochondrial ATP fluorescence probe was used to assess mitochondrial function.Cytoskeletal remodeling was detected with a microfilament green fluorescent probe(Actin-Tracker Green-488).Transwell assay was used to assess cell invasion ability.The expression levels of ATP5J and translocase of outer mitochondrial membrane 20(TOMM20)were determined by Western blot.RESULTS:Overexpression of ATP5J up-regulated mitochondrial membrane potential and mitochondrial ATP fluorescence intensity,induced cytoskeletal remodeling,promoted cell invasion and TOMM20 expression,and inhibited ROS production(P<0.01).On the contrary,knockdown of ATP5J significantly decreased mitochondrial membrane potential and mitochondrial ATP fluorescence intensity,significantly decreased cell invasion ability and TOMM20 expression,promoted ROS production and blocked cytoskeletal remodeling(P<0.01).CONCLUSION:ATP5J regulates mitochondrial energy transformation in hepatocellular carcinoma cells,and affects metastasis of hepatoma carcinoma cells by regulating mitochondrial membrane potential and mitochondrial ATP production-mediated cytoskeletal remodeling through TOMM20.

关 键 词：肝细胞癌 线粒体功能 ATP5J蛋白 细胞骨架 

分 类 号：R735.7[医药卫生—肿瘤] R363.2[医药卫生—临床医学] R575.5