基质刚度对肝癌细胞增殖及糖代谢的影响
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重庆大学

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Q66

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Effect of matrix stiffness on proliferation and glucose metabolism of hepatocellular carcinoma cells
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Chongqing University

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    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肝癌细胞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基质刚度上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-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose,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细胞的增殖能力、葡萄糖摄取和Glut1的表达都显著增加。糖酵解途径被阻断后,不同基质刚度上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;本研究为肝癌的临床治疗以及糖酵解代谢为治疗靶点的药物开发提供了相应的实验依据。

    Abstract:

    Objective To investigate the proliferation ability and glucose metabolism of hepatocellular carcinoma (HCC) cell growth on different stiffness of matrix and to explore relevance between metabolism and biological behavior variety of HCC cells caused by matrix stiffness.Methods The influence of different stiffness of matrix on proliferation of HepG2 cell was detected by CCK-8 assay and cell count assay.2-NBDG and Flow cytometry was 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 HepG2 cells proliferation was detected. Result The proliferation ability, glucose uptake and the expression of Glut1 of HepG2 cell increased with increasing stiffness of matrix. When glycolysis was inhibited, the proliferation ability of HepG2 cell growth on different stiffness of matrix was similar. Conclusions The mechanical microenvironment has an important effect on HCC cells proliferation; stiff matrix may promote HCC cells proliferation through increasing glycolysis; this study provides the experimental basis for the clinical treatment of hepatocellular carcinoma and drug development targeting glucose metabolism.

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  • 收稿日期:2019-01-15
  • 最后修改日期:2019-02-11
  • 录用日期:2019-02-18
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