Yang Yuping, Liu Lingling, Sun Jinghui, Wang Shu, Yang Zhongyuan, Li Honghui, Huang Na, Zhao Wei
Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Chengdu Medical College, Chengdu, China.
School of Laboratory Medicine/Sichuan Provincial Engineering Laboratory for Prevention and Control Technology of Veterinary Drug Residue in Animal-origin Food, Chengdu Medical College, Chengdu, China.
Front Oncol. 2021 Feb 24;11:629543. doi: 10.3389/fonc.2021.629543. eCollection 2021.
Cancer cell proliferation is a metabolically demanding process that requires high rate of glycolysis to support anabolic growth. Deoxypodophyllotoxin (DPT) is a natural flavonolignan with various pharmacological activities, including antitumor effect. However, whether DPT affects the metabolic reprogramming of cancer cells is unknown. The purpose of this study is to investigate the role of DPT on non-small cell lung cancer (NSCLC) and to explore whether HIF-1α-mediated glycolysis is involved in its mechanism of action.The level of HIF-1α mRNA and protein in NSCLC cells following DPT treatment was detected using qRT-PCR and western blotting, respectively. Cell Counting Kit-8 (CCK-8) and caspase-3 activity assays were performed to analyze cell proliferation and apoptosis. The underlying molecular mechanism was identified by dual luciferase assay, Western blotting, qRT-PCR, glucose consumption, lactate production, and immunoprecipitation. A murine NSCLC model was used to clarify the effect of DPT treatment on tumor cell proliferation. Our findings showed that DPT treatment inhibited NSCLC cell growth in a dose- and time-dependent manner. Further analysis suggested that DPT treatment inhibited HIF-1α signaling pathway by Parkin-mediated protein degradation in NSCLC cells. DPT treatment significantly decreased glucose consumption and lactate production. In addition, DPT treatment reduced the expression of HIF-1α target genes, including GLUT1, HK2 and LDHA, resulting in reduction in glycolysis. We further revealed that DPT-induced cell growth inhibition and increased glucose and lactate levels could be reversed by overexpressing HIF-1α. Additionally, we found that DPT repressed NSCLC growth and GLUT1, HK2 and LDHA expression . Overall, this study suggested that DPT inhibited NSCLC growth by preventing HIF-1α-mediated glycolysis.
癌细胞增殖是一个代谢需求很高的过程，需要高速糖酵解来支持合成代谢生长。脱氧鬼臼毒素（DPT）是一种具有多种药理活性的天然黄酮木脂素，包括抗肿瘤作用。然而，DPT是否影响癌细胞的代谢重编程尚不清楚。本研究的目的是探讨DPT在非小细胞肺癌（NSCLC）中的作用，并探讨HIF-1α介导的糖酵解是否参与其作用机制。分别采用qRT-PCR和蛋白质免疫印迹法检测DPT处理后NSCLC细胞中HIF-1α mRNA和蛋白水平。进行细胞计数试剂盒-8（CCK-8）和caspase-3活性检测以分析细胞增殖和凋亡。通过双荧光素酶检测、蛋白质免疫印迹法、qRT-PCR、葡萄糖消耗、乳酸生成和免疫沉淀确定潜在的分子机制。使用小鼠NSCLC模型阐明DPT处理对肿瘤细胞增殖的影响。我们的研究结果表明，DPT处理以剂量和时间依赖性方式抑制NSCLC细胞生长。进一步分析表明，DPT处理通过帕金森蛋白介导的蛋白质降解抑制NSCLC细胞中的HIF-1α信号通路。DPT处理显著降低葡萄糖消耗和乳酸生成。此外，DPT处理降低了HIF-1α靶基因的表达，包括葡萄糖转运蛋白1（GLUT1）、己糖激酶2（HK2）和乳酸脱氢酶A（LDHA），导致糖酵解减少。我们进一步发现，过表达HIF-1α可以逆转DPT诱导的细胞生长抑制以及葡萄糖和乳酸水平的升高。此外，我们发现DPT抑制NSCLC生长以及GLUT1、HK2和LDHA的表达。总体而言，本研究表明DPT通过阻止HIF-1α介导的糖酵解来抑制NSCLC生长。
