摘要
目的研究不同基质刚度对肝癌细胞HepG2增殖和糖代谢的影响,探究细胞外基质(extracellular matrix,ECM)刚度对肝癌细胞代谢和生物学行为影响的相关性。方法采用CCK8和细胞计数法检测培养在不同刚度基质上HepG2细胞增殖的变化,利用2-(N-7-硝基-2,1,3-苯并恶二唑-4-氨基)-2-脱氧-D-葡萄糖(2-NBDG)并通过流式细胞术检测基质刚度对HepG2葡萄糖摄取的影响,利用荧光定量RT-PCR检测基质刚度对HepG2葡萄糖转运蛋白1(glucose transporter 1,Glut1)表达的影响,并利用糖酵解抑制剂2-脱氧葡萄糖(2-deoxy-D-glucose,2-DG)阻断糖酵解途径后检测基质刚度对HepG2细胞增殖的影响。结果随基质刚度增加,HepG2细胞增殖能力、葡萄糖摄取和Glut1表达都显著增加。糖酵解途径被阻断后,不同刚度基质上HepG2细胞增殖能力基本相同。结论肝癌细胞所处力学微环境对肝癌细胞的增殖有重要影响;较大的基质刚度可能通过调控糖代谢途径促进肝癌细胞的增殖。研究结果为肝癌的临床治疗以及糖代谢为治疗靶点的药物开发提供相应的实验依据。
Objective To investigate the influences of different matrix stiffness on proliferation ability and glucose metabolism of hepatocellular carcinoma(HCC)cells and to explore the correlation between metabolism and biological behavior changes of HCC cells resulted from the stiffness of extracellular matrix(ECM).Methods The proliferation changes of HepG2 cells cultured on matrix with different stiffness were detected by CCK-8 assay and cell count assay.2-NBDG and flow cytometry were used to detect the effect of matrix stiffness on glucose uptake.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression level of Glut1.Then,2-DG was used to inhibit glycolysis,and the influences of matrix stiffness on proliferation of HepG2 cells were detected.Results The proliferation ability,glucose uptake and the expression of Glut1 of HepG2 cells increased with the matrix stiffness increasing.When glycolysis was inhibited,the proliferation ability of HepG2 cells grown on matrix with different stiffness was similar.Conclusions The mechanical microenvironment had an important effect on proliferation of HCC cells;matrix with a larger stiffness might promote proliferation of HCC cells through regulating glycolysis.The research findings provide a corresponding experimental basis for the clinical treatment of HCC cells and drug development targeting glucose metabolism.
作者
刘秋萍
田博仁
罗庆
宋关斌
LIU Qiuping;TIAN Boren;LUO Qing;SONG Guanbin(College of Bioengineering,Chongqing University,Chongqing 400030,China)
出处
《医用生物力学》
EI
CAS
CSCD
北大核心
2019年第2期133-138,共6页
Journal of Medical Biomechanics
基金
国家自然科学基金项目(11832008)
关键词
基质刚度
肝癌细胞
细胞增殖
糖代谢
生物力学
extracellular matrix(ECM)
hepatocellular carcinoma(HCC)cell
glucose metabolism
cell proliferation
biomechanics
作者简介
通信作者:宋关斌,教授,博士研究生导生,E-mail:song@cqu.edu.cn
