艾地苯醌通过调控线粒体活性氧抑制肝癌生长及转移的作用及机制  

Idebenone inhibiting growth and metastasis of hepatocellular carcinoma via regulating mitochondrial reactive oxygen species

作　　者：唐津天[1] 唐润娟[2] 薛峰[1] 易超[1] 黎旺红 刘晨 TANG Jintian;TANG Runjuan;XUE Feng;YI Chao;LI Wanghong;LIU Chen(Department of Hepatobiliary and Pancreatic Surgery,Cancer Hospital Af filiated to Xinjiang Medical University,Urumqi,Xinjiang 830000,China;Department of Rehabilitation,Second Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830000,China)

机构地区：[1]新疆医科大学附属肿瘤医院肝胆胰外科,新疆乌鲁木齐830000 [2]新疆医科大学第二附属医院康复科,新疆乌鲁木齐830000

出　　处：《中华肿瘤防治杂志》2024年第6期354-360,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：新疆维吾尔自治区自然科学基金(2021D01C399)。

摘　　要：目的 探究艾地苯醌(IDE)通过调控线粒体活性氧(ROS)抑制肝癌生长和转移的功能及其机制。方法 细胞计数盒8(CCK8)法检测不同浓度(1、2、4、6、8和16μmol/L浓度孵育24 h)和不同时间(10μmol/L孵育6、12、24、48和72 h)IDE处理后肝癌细胞HepG2的细胞活力影响。将10μmol/L IDE处理的HepG2细胞分为IDE-R组和IDE-T2组,将二甲基亚砜(DMSO)处理的分为DMSO-R组和DMSO-T2组。Transwell实验分析IDE-T2和DMSO-T2组细胞侵袭和迁移,2,7-二氯二氢荧光素二乙酸酯(DCFHDA)和MitoSOX探针分别检测IDE-R和DMSO-R组细胞总体ROS和线粒体ROS水平。使用HepG2肝癌原位移植裸鼠模型,将其分为IDE-L组和DMSO-L组(8只/组),组织病理学法检测肿瘤的肺转移能力;使用HepG2细胞构建肝癌皮下瘤模型,将其分为IDE-M组和DMSO-M组(8只/组),检测肿瘤生长大小。结果 IDE可以有效抑制肝癌细胞HepG2的细胞活力,1、2、4、6、8和16μmol/L IDE处理24 h细胞抑制率分别为(3.33±0.88)%、(9.67±1.20)%、(16.31±1.15)%、(37.36±2.41)%、(61.42±1.76)%和(86.71±2.02)%,24 h的半数抑制浓度(IC_(50))为(8.09±0.28)μmol/L;10μmol/L IDE处理HepG2细胞6、12、24、48和72 h的细胞抑制率分别为(11.04±1.52)%、(31.35±1.76)%、(51.37±1.53)%、(75.38±2.37)%和(90.56±1.93)%。细胞内ROS检测发现,IDE-R组线粒体ROS水平[(12 017.34±1 647.23)vs(57 495.67±2 512.15)]和细胞总ROS水平[(11 411.57±1 119.49)vs(51 132.26±2 018.33)]均低于DMSO-R组,差异有统计学意义,t值分别为-26.30和-29.81,P值分别为<0.001和0.009。Transwell实验结果显示,IDE-T2组[(27.35±3.14)%,(22.19±2.54)%]细胞侵袭和迁移能力均低于DMSO-T2组[(100.00±2.08)%,(100.00±3.13)%]。体内肿瘤转移分析发现,与DMSO-L组(73.12±3.65)相比,IDE-L组(23.67±2.39)裸鼠肺转移的数量降低,差异有统计学意义,t=18.56,P<0.001。皮下成瘤实验显示,IDE-M组肿瘤块体积较DMSO-M组小。结论 抗氧化剂IDE可以通过清除线粒体ROS从而抑制肝癌�Objective To investigate the function and mechanism of Idebenone(IDE)in inhibiting hepatocellular carcinoma metastasis by regulating mitochondrial reactive oxygen species(ROS).Methods Cell counting kit 8(CCK8)method was used to detect the cell viability of HepG2 liver cancer cells at different concentrations(incubation for 24 hours at 1,2,4,6,8 and 16μmol/L IDE)and at different time(incubation with 10μmol/L IDE for 6,12,24,48 and 72 h).HepG2 cells treated with 10μmol/L IDE were divided into IDE-R group and IDE-T2 group.HepG2 cells treated with dimethyl sulfoxide(DMSO)were divided into DMSO-R group and DMSO-T2 group.Transwell experiment was used to analyze cell invasion and migration in IDE-T2 and DMSO-T2 groups.2,7-Dichlorodihydrofluorescein diacetate(DCFH-DA)and MitoSOX probes were used to detect overall ROS and mitochondrial ROS levels in IDE-R and DMSO-R groups.HepG2 hepatocellular carcinoma orthotopic transplantation mouse models were divided into IDE-L group and DMSO-L group(8 mice in each group),and the lung metastasis ability of the tumors was detected by histopathology.HepG2 cells were used to construct subcutaneous tumor model of hepatocellular carcinoma,which was divided into IDE-M group and DMSO-M group,8 cells in each group,and the tumor growth size was detected.Results IDE could effectively inhibit the HepG2 cell viability.The cell inhibition rates at concentrations of 1,2,4,6,8 and 16μmol/L IDE treatment for 24 hours were(3.33±0.88)%,(9.67±1.20)%,(16.31±1.15)%,(37.36±2.41)%,(61.42±1.76)%and(86.71±2.02)%,respectively.The50%inhibitory concentration(IC_(50))at 24 hours was 8.09±0.28μmol/L.The cell inhibition rates of HepG2 cells treated with 10μmol/LIDEfor 6,12,24,48 and 72 hours were(11.04±1.52)%,(31.35±1.76)%,(51.37±1.53)%,(75.38±2.37)%and(90.56±1.93)%,respectively.Intracellular ROS detection revealed that the mitochondrial ROS levels in the IDE-R group(12017.34±1647.23)were lower than those in the DMSO-R group(57495.67±2512.15),with a statistically significant difference,t=-26.30

关 键 词：肝癌 转移 艾地苯醌 活性氧 线粒体 

分 类 号：R735.7[医药卫生—肿瘤]