Isayev Orkhan, Rausch Vanessa, Bauer Nathalie, Liu Li, Fan Pei, Zhang Yiyao, Gladkich Jury, Nwaeburu Clifford C, Mattern Jürgen, Mollenhauer Martin, Rückert Felix, Zach Sebastian, Haberkorn Uwe, Gross Wolfgang, Schönsiegel Frank, Bazhin Alexandr V, Herr Ingrid
Molecular OncoSurgery, University of Heidelberg and German Cancer Research Center (DKFZ), Heidelberg, Germany; General and Transplantation Surgery, University Hospital Heidelberg, Germany.
Oncotarget. 2014 Jul 15;5(13):5177-89. doi: 10.18632/oncotarget.2120.
According to the cancer stem cell (CSC) hypothesis, the aggressive growth and early metastasis of pancreatic ductal adenocarcinoma (PDA) is due to the activity of CSCs, which are not targeted by current therapies. Otto Warburg suggested that the growth of cancer cells is driven by a high glucose metabolism. Here, we investigated whether glycolysis inhibition targets CSCs and thus may enhance therapeutic efficacy. Four established and 3 primary PDA cell lines, non-malignant cells, and 3 patient-tumor-derived CSC-enriched spheroidal cultures were analyzed by glucose turnover measurements, MTT and ATP assays, flow cytometry of ALDH1 activity and annexin positivity, colony and spheroid formation, western blotting, electrophoretic mobility shift assay, xenotransplantation, and immunohistochemistry. The effect of siRNA-mediated inhibition of LDH-A and LDH-B was also investigated. The PDA cells exhibited a high glucose metabolism, and glucose withdrawal or LDH inhibition by siRNA prevented growth and colony formation. Treatment with the anti-glycolytic agent 3-bromopyruvate almost completely blocked cell viability, self-renewal potential, NF-κB binding activity, and stem cell-related signaling and reverted gemcitabine resistance. 3-bromopyruvate was less effective in weakly malignant PDA cells and did not affect non-malignant cells, predicting minimal side effects. 3-bromopyruvate inhibited in vivo tumor engraftment and growth on chicken eggs and mice and enhanced the efficacy of gemcitabine by influencing the expression of markers of proliferation, apoptosis, self-renewal, and metastasis. Most importantly, primary CSC-enriched spheroidal cultures were eliminated by 3-bromopyruvate. These findings propose that CSCs may be specifically dependent on a high glucose turnover and suggest 3-bromopyruvate for therapeutic intervention.
根据癌症干细胞（CSC）假说，胰腺导管腺癌（PDA）的侵袭性生长和早期转移归因于CSC的活性，而目前的治疗方法无法靶向这些细胞。奥托·瓦尔堡提出癌细胞的生长由高糖代谢驱动。在此，我们研究了糖酵解抑制是否靶向CSC，从而可能提高治疗效果。通过葡萄糖周转率测量、MTT和ATP测定、ALDH1活性和膜联蛋白阳性的流式细胞术、集落和球体形成、蛋白质印迹、电泳迁移率变动分析、异种移植和免疫组织化学，对4种已建立的和3种原发性PDA细胞系、非恶性细胞以及3种患者肿瘤来源的富含CSC的球体培养物进行了分析。还研究了siRNA介导的LDH-A和LDH-B抑制的效果。PDA细胞表现出高糖代谢，葡萄糖剥夺或siRNA介导的LDH抑制可阻止细胞生长和集落形成。用抗糖酵解剂3-溴丙酮酸处理几乎完全阻断了细胞活力、自我更新潜力、NF-κB结合活性以及干细胞相关信号传导，并逆转了吉西他滨耐药性。3-溴丙酮酸在低恶性PDA细胞中的效果较差，且不影响非恶性细胞，预示着副作用最小。3-溴丙酮酸抑制了鸡胚和小鼠体内肿瘤的植入和生长，并通过影响增殖、凋亡、自我更新和转移标志物的表达增强了吉西他滨的疗效。最重要的是，3-溴丙酮酸消除了原发性富含CSC的球体培养物。这些发现表明CSC可能特别依赖高葡萄糖周转率，并提示3-溴丙酮酸可用于治疗干预。
