CT-707对缺氧微环境下肝癌细胞能量代谢和细胞周期的影响及作用机制  

Effect of CT-707 on Energy Metabolism and Cell Cycle of Hepatocellular Carcinoma Cells in Hypoxic Microenvironment and Its Mechanism

作　　者：姚红兵 吴嘉兴 郭威 文雪霖 蒋建晖 刘翔峰[2] Yao Hongbing;Wu Jiaxing;Guo Wei(Department of Hepatobiliary Surgery,The Second Affiliated Hospital of Guilin Medical University,Guangxi 541199,China)

机构地区：[1]桂林医学院第二附属医院肝胆外科,541199 [2]中南大学湘雅二医院急诊医学科,长沙410011

出　　处：《医学研究杂志》2021年第11期69-75,共7页Journal of Medical Research

基　　金：广西壮族自治区自然科学基金资助项目(面上项目)(2018GXNSFAA050094)。

摘　　要：目的研究CT-707对缺氧微环境下肝癌细胞能量代谢和细胞周期的影响及作用机制。方法HepG2细胞分为对照组和CT-707高(6μmol/L)、中(3μmol/L)、低(1.5μmol/L)剂量组,缺氧(2%O_(2))条件下培养,采用CCK8法测定细胞活力,HK及LDH试剂盒检测细胞中HK酶活力及LDH含量,Western blot法检测cyclinD1、cyclin E、p-FAK/FAK、p-PI_(3)K/PI_(3)K、p-Akt/Akt、p-GSK-3β/GSK-3β蛋白表达。构建HepG2细胞裸鼠皮下成瘤模型,分为模型组和CT-707高(100 mg/kg)、中(50mg/kg)、低(25mg/kg)剂量组,每组8只,观察并测量肿瘤质量及体积,采用HE染色进行瘤组织病理学观察,免疫组化法检测肿瘤组织中PCNA的表达。结果与对照组比较,CT-707各剂量组均可显著降低HepG2细胞活力、HK活性及LDH含量,显著下调cyclin D1、cyclin E、p-FAK/FAK、p-PI_(3)K/PI_(3)K、p-Akt/Akt、p-GSK-3β/GSK-3β蛋白表达水平(P<0.05),中、高剂量组的效应明显优于低剂量组(P<0.05),中、高剂量组间结果比较,差异无统计学意义(P>0.05)。与模型组比较,CT-707各剂量组均可显著减小瘤质量和瘤体积,降低PCNA的蛋白表达水平(P<0.05),中、高剂量组的效应明显优于低剂量组(P<0.05),中、高剂量组间结果比较差异无统计学意义(P>0.05)。结论CT-707可抑制缺氧环境下肝癌细胞能量代谢,进而诱导细胞周期阻滞,抑制肿瘤生长,其机制可能与抑制PI 3K/Akt/GSK-3β信号通路激活相关。Objective To investigate the effect and mechanism of CT-707 on energy metabolism and cell cycle of hepatocellular carcinoma cells under hypoxic microenvironment.Methods HepG2 cells were divided into control group and CT-707 high(6μmol/L),medium(3μmol/L)and low(1.5μmol/L)dose groups,which were cultured under hypoxia(2%O 2).The cell viability was determined by CCK8 assay,and the HK enzyme activity and LDH content in the cells were detected by HK kit and LDH kit respectively.The expressions of cyclin D1,cyclin E,p-FAk/FAK,p-PI 3K/PI 3K、p-Akt/Akt and p-GSK-3β/GSK-3βwere evaluated by Western blot.HepG2 cells were used to establish subcutaneous tumor models of nude mice,which was divided into model group and CT-707 high(100mg/kg),medium(50mg/kg)and low(25mg/kg)dose groups and with 8 mice in each group,then tumor size and volume were observed and measured.HE staining was used for tumor histopathological observation,and immunohistochemistry was used to detect the expression of PCNA in tumor tissues.Results Compared with the control group,the cell vitality,HK enzyme activity and LDH content were significantly reduced,and the expression levels of cyclin D1,cyclin E,p-FAK/FAK,p-PI 3K/PI 3K,p-Akt/Akt and p-GSK-3β/GSK-3βwere down-regulatedmarkedly in HepG2 cells of each dose of CT-707(P<0.05).The effect of high and medium dose groups was obviously better than the low dose group(P<0.05),and there was no statistical difference between medium and high dose group(P>0.05).Compared with the model group,the tumor mass,tumor volume and the expression level of PCNA protein were significantly reduced in each dose of CT-707(P<0.05).The effect of the medium and high dose groups was significantly better than low dose group(P<0.05),and there was no statistical difference between the medium and high dose groups(P>0.05).Conclusion CT-707 can inhibit the energy metabolism of hepatocellular carcinoma cells in hypoxic microenvironment and induce cell cycle arrest,which is closely related to inhibiting the activation of PI 3K/Akt/GSK-3�

关 键 词：CT-707 缺氧 肝癌 能量代谢 细胞周期 

分 类 号：R575[医药卫生—消化系统]