一种新型热休克蛋白抑制剂 KU757 在 lenvatinib 耐药滤泡甲状腺癌细胞中具有疗效，可克服耐药细胞中上调的糖酵解作用。
A novel heat shock protein inhibitor KU757 with efficacy in lenvatinib-resistant follicular thyroid cancer cells overcomes up-regulated glycolysis in drug-resistant cells in vitro.
机构信息
Department of Surgery, University of Michigan, Ann Arbor, MI.
Department of Pharmacology, University of Michigan, Ann Arbor, MI.
出版信息
Surgery. 2021 Jan;169(1):34-42. doi: 10.1016/j.surg.2020.06.009. Epub 2020 Jul 24.
BACKGROUND
Patients with advanced differentiated thyroid cancer develop resistance to lenvatinib treatment from metabolic dysregulation. Heat shock protein 90 is a molecular chaperone that plays an important role in glycolysis and metabolic pathway regulation. We hypothesize that lenvatinib-resistant differentiated thyroid cancer cells will have an increased dependency on glycolysis and that a novel C-terminal heat shock protein 90 inhibitor (KU757) can effectively treat lenvatinib-resistant cells by targeting glycolysis.
METHODS
Inhibitory concentration 50 values of thyroid cancer cells were determined by CellTiter-Glo assay (Promega Corp, Madison, WI). Glycolysis was measured through Seahorse experiments. Reverse transcription-polymerase chain reaction and Western blot evaluated glycolytic pathway genes/proteins. Exosomes were isolated/validated by nanoparticle tracking analysis and Western blot. Differentially expressed long non-coding ribonucleic acids in exosomes and cells were evaluated using quantitative polymerase chain reaction.
RESULTS
Extracellular acidification rate demonstrated >2-fold upregulation of glycolysis in lenvatinib-resistant cells versus parent cells and was downregulated after KU757 treatment. Lenvatinib-resistant cells showed increased expression of the glycolytic genes lactic acid dehydrogenase, pyruvate kinase M1/2, and hexokinase 2. KU757 treatment resulted in downregulation of these genes and proteins. Several long non-coding ribonucleic acids associated with glycolysis were significantly upregulated in WRO-lenvatinib-resistant cells and exosomes and downregulated after KU757 treatment.
CONCLUSION
Lenvatinib resistance leads to increased glycolysis, and KU757 effectively treats lenvatinib-resistant cells and overcomes this increased glycolysis by targeting key glycolytic genes, proteins, and long non-coding ribonucleic acids.
背景
晚期分化型甲状腺癌患者因代谢失调对仑伐替尼治疗产生耐药性。热休克蛋白 90 是一种分子伴侣，在糖酵解和代谢途径调节中发挥重要作用。我们假设仑伐替尼耐药的分化型甲状腺癌细胞将对糖酵解有更高的依赖性，并且新型 C 端热休克蛋白 90 抑制剂（KU757）可以通过靶向糖酵解有效地治疗仑伐替尼耐药细胞。
方法
通过 CellTiter-Glo 测定法（Promega 公司，麦迪逊，威斯康星州）测定甲状腺癌细胞的半数抑制浓度 50 值。通过 Seahorse 实验测量糖酵解。逆转录-聚合酶链反应和 Western blot 评估糖酵解途径基因/蛋白。通过纳米颗粒跟踪分析和 Western blot 分离/验证外泌体。使用定量聚合酶链反应评估外泌体和细胞中差异表达的长链非编码核糖核酸。
结果
细胞外酸化率显示，与亲本细胞相比，仑伐替尼耐药细胞的糖酵解上调了>2 倍，并且在 KU757 处理后下调。仑伐替尼耐药细胞表现出糖酵解基因乳酸脱氢酶、丙酮酸激酶 M1/2 和己糖激酶 2 的表达增加。KU757 处理导致这些基因和蛋白的下调。与糖酵解相关的几种长链非编码核糖核酸在 WRO-仑伐替尼耐药细胞和外泌体中显著上调，并在 KU757 处理后下调。
结论
仑伐替尼耐药导致糖酵解增加，KU757 通过靶向关键糖酵解基因、蛋白和长链非编码核糖核酸有效治疗仑伐替尼耐药细胞并克服这种增加的糖酵解。
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