Ren Ying, Gu Yue Kai, Li Zhen, Xu Guang Zi, Zhang Yang Meng, Dong Min Xin, Wang Ying, Zhou Xi Bing
Department of Pathology, Henan Province People's Hospital, Zhengzhou University People's Hospital Zhengzhou, Henan, China.
Henan University Kaifeng, Henan, China.
Am J Transl Res. 2020 Mar 15;12(3):825-836. eCollection 2020.
Currently, the acquired resistance of the hepatocellular carcinoma (HCC) first-line therapeutic agent-sorafenib (SOR) remains a major challenge for HCC management. Recent evidence suggested the association between CXCL/CXCRs chemokines and chemotherapy resistant in cancer cells. Hence, exploring the internal mechanism of CXCRs involved in SOR resistance will help to improve the efficacy of HCC. SOR-resistant HCC cells (Huh7-SOR) were established through escalating concentration of SOR. Glucose consumption, lactate production, intracellular ATP levels and oxygen consumption of HCC cells were determined by using the associated detected kits. Effects of CXCR3 on metabolic phenotype of HCC cells, AMPK pathway activity and adipocytokines were demonstrated by knocking down CXCR3 expression with the CXCR3 siRNA technique combined with qPCR and western blot. During the indicated procedure, SOR-resistant HCC cells-Huh7-SOR presented EMT-like morphologic change and underwent glycolysis to OXPHOS switch, representing reduced glucose consumption and lactate production, but increased oxygen consumption level and intercellular ATP levels. Moreover, metabolic alteration in SOR-resistance HCC cells was mediated by CXCR3. Mechanistically, CXCR3 induced metabolic alteration in SOR-resistance HCC cells through downregulating AMPK pathway activity and lipid peroxidation as well as upregulating levels of adipocytokines. The activation of A MPK pathway with metformin achieved the sensitization of HCC to SOR treatment in vivo. These findings unravel the association between metabolic alteration and SOR-resistance in HCC cells and demonstrate an important role of CXCR3 in the development of HCC cells resistance to SOR treatment and a novel mechanism of CXCR3 regulating AMPK pathway activity and adipocytokine signaling, lipid peroxidation resulted in metabolic alteration during the chemoresistance.
目前,肝细胞癌(HCC)一线治疗药物索拉非尼(SOR)的获得性耐药仍然是HCC治疗的主要挑战。最近的证据表明CXCL/CXCRs趋化因子与癌细胞化疗耐药之间存在关联。因此,探索CXCRs参与SOR耐药的内在机制将有助于提高HCC的疗效。通过逐步提高SOR浓度建立了SOR耐药的HCC细胞(Huh7-SOR)。使用相关检测试剂盒测定HCC细胞的葡萄糖消耗、乳酸生成、细胞内ATP水平和耗氧量。通过CXCR3 siRNA技术结合qPCR和蛋白质印迹法敲低CXCR3表达,证明了CXCR3对HCC细胞代谢表型、AMPK途径活性和脂肪细胞因子的影响。在所示过程中,SOR耐药的HCC细胞-Huh7-SOR呈现出类似上皮-间质转化(EMT)的形态变化,并经历了从糖酵解到氧化磷酸化的转变,表现为葡萄糖消耗和乳酸生成减少,但耗氧量水平和细胞间ATP水平增加。此外,SOR耐药HCC细胞中的代谢改变由CXCR3介导。机制上,CXCR3通过下调AMPK途径活性和脂质过氧化以及上调脂肪细胞因子水平,诱导SOR耐药HCC细胞发生代谢改变。用二甲双胍激活AMPK途径可在体内实现HCC对SOR治疗的致敏。这些发现揭示了HCC细胞中代谢改变与SOR耐药之间的关联,并证明了CXCR3在HCC细胞对SOR治疗耐药发展中的重要作用,以及CXCR3调节AMPK途径活性和脂肪细胞因子信号传导的新机制,脂质过氧化在化疗耐药期间导致代谢改变。