Enhancing 5-fluorouracil efficacy through suppression of PKM2 in colorectal cancer cells
Enhancing 5-fluorouracil efficacy through suppression of PKM2 in colorectal cancer cells
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DOI：
10.1007/s00280-018-3676-7
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发表时间：
2018-12-01
影响因子：
3
通讯作者：
Zheng, Changqing
中科院分区：
文献类型：
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作者：
Cao, Yong;Lin, Yan;Zheng, Changqing
目的癌细胞改变常规代谢途径以维持快速增殖。癌组织中代谢重塑的一个例子是丙酮酸激酶同工酶 M2 (PKM2) 的上调，该酶参与有氧糖酵解。事实上，PKM2 此前已被确定为肿瘤生物标志物和癌症治疗的潜在靶点。本研究评估了 PKM2 在 5-氟尿嘧啶 (5-FU) 结直肠癌 (CRC) 抗癌功效中的作用。方法通过转染表达针对 PKM2 的短发夹 RNA (shRNA) 的慢病毒载体，使用 HCT116、SW480 和 HT-29 细胞。针对 5-FU 处理，检查细胞增殖，监测 ATP/ADP 比值水平，测量 5-FU 细胞内积累，并使用液相色谱-质谱 (LC-MS) 评估细胞内磷酸烯醇丙酮酸 (PEP)、丙酮酸和乳酸水平。通过CRC皮下肿瘤模型研究抑制PKM2对5-FU体内疗效的影响。结果抑制PKM2导致葡萄糖代谢变化，导致三磷酸腺苷(ATP)合成减少。 ATP/ADP 比率水平降低导致 5-FU 在细胞内积聚，从而增强该药物在多种 CRC 细胞系中的治疗效果。此外，在体内 HCT116 CRC 模型中证明了 5-FU 通过同时抑制 PKM2 来增强疗效。结论我们表明，与单独使用 5-FU 相比，联合治疗显示出更优异的抗癌功效。这些发现表明，靶向 PKM2 可以提高化疗的疗效，可能为改善 CRC 患者的化疗结果提供一种新方法。
PurposeCancer cells alter regular metabolic pathways in order to sustain rapid proliferation. One example of metabolic remodeling in cancerous tissue is the upregulation of pyruvate kinase isoenzyme M2 (PKM2), which is involved in aerobic glycolysis. Indeed, PKM2 has previously been identified as a tumor biomarker and as a potential target for cancer therapy. Here, the role of PKM2 in the anticancer efficacy of 5-fluorouracil (5-FU) was evaluated in colorectal cancer (CRC).MethodsHCT116, SW480 and HT-29 cells were used by transfection with lentiviral vectors expressing short hairpin RNA (shRNA) against PKM2. In response to 5-FU treatment, cellular proliferation was examined, the levels of ATP/ADP ratio were monitored, the intracellular accumulation of 5-FU was measured, and intracellular levels of phosphoenolpyruvate (PEP), pyruvate and lactate were evaluated by using liquid chromatography-mass spectrometry (LC-MS). A CRC subcutaneous tumor model was performed to investigate the effect of PKM2 inhibition on 5-FU efficacy in vivo.ResultsSuppression of PKM2 resulted in changes in glucose metabolism, leading to decreased synthesis of adenosine triphosphate (ATP). Reduced levels of ATP/ADP ratio resulted in the intracellular accumulation of 5-FU, consequently enhancing the therapeutic efficacy of this drug in several CRC cell lines. Furthermore, the enhanced efficacy of 5-FU by simultaneous inhibition of PKM2 was demonstrated in an in vivo HCT116 CRC model.ConclusionWe show that the combination treatment showed superior anticancer efficacy as compared to 5-FU alone. These findings suggest that targeting PKM2 can increase the efficacy of chemotherapy, potentially providing a new approach for improving the outcome of chemotherapy in patients with CRC.