Butyrate Suppresses the Proliferation of Colorectal Cancer Cells via Targeting Pyruvate Kinase M2 and Metabolic Reprogramming
Butyrate Suppresses the Proliferation of Colorectal Cancer Cells via Targeting Pyruvate Kinase M2 and Metabolic Reprogramming
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丁酸盐通过靶向丙酮酸激酶 M2 和代谢重编程抑制结直肠癌细胞的增殖
DOI：
10.1074/mcp.ra118.000752
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发表时间：
2018-08-01
影响因子：
7
通讯作者：
Hao, Haiping
中科院分区：
文献类型：
--
作者：
Li, Qingran;Cao, Lijuan;Hao, Haiping
丁酸盐是一种短链脂肪酸，在肠腔中以高浓度存在。已充分证明丁酸盐通过充当高能代谢物促进正常结肠细胞的增殖，同时通过充当组蛋白脱乙酰酶抑制剂，表观遗传地抑制经历瓦尔堡效应的癌对应物的增殖。然而，丁酸盐如何中断结直肠癌细胞的代谢并最终导致细胞增殖受到抑制仍不清楚。在这里，我们采用代谢组学-蛋白质组学相结合的方法来探索丁酸盐介导的增殖停滞和细胞代谢之间的联系。一项代谢组学研究显示，丁酸盐处理的HCT-116细胞的代谢谱发生了重塑，丙酮酸盐显著蓄积，丙酮酸盐上游糖酵解中间产物减少，核苷酸水平降低。补充关键代谢物中间体直接影响癌细胞代谢，并调节HCT-116细胞中丁酸盐的抑制作用。通过基于药物亲和反应靶点稳定性（DARTS）的定量蛋白质组学方法，我们揭示了丙酮酸激酶PKM 2的M2亚型作为丁酸盐的直接结合靶点。丁酸盐通过促进PKM 2的去磷酸化和四聚化激活PKM 2，从而重新编程结直肠癌细胞的代谢，抑制瓦尔堡效应，同时有利于能量代谢。因此，我们的研究提供了PKM 2诱导的代谢重塑和丁酸盐的抗肿瘤功能之间的机制联系，并证明了一种广泛适用的方法来揭示具有生物学功能的小分子的未知蛋白质靶点。
Butyrate is a short chain fatty acid present in a high concentration in the gut lumen. It has been well documented that butyrate, by serving as an energetic metabolite, promotes the proliferation of normal colonocytes while, by serving as a histone deacetylase inhibitor, epigenetically suppressing the proliferation of cancerous counterparts undergoing the Warburg effect. However, how butyrate interrupts the metabolism of colorectal cancer cells and ultimately leads to the suppression of cell proliferation remains unclear. Here, we employed a metabolomics-proteomics combined approach to explore the link between butyrate-mediated proliferation arrest and cell metabolism. A metabolomics study revealed a remodeled metabolic profile with pronounced accumulation of pyruvate, decreased glycolytic intermediates upstream of pyruvate and reduced levels of nucleotides in butyrate-treated HCT-116 cells. Supplementation of key metabolite intermediates directly affected cancer-cell metabolism and modulated the suppressive effect of butyrate in HCT-116 cells. By a Drug Affinity Responsive Target Stability (DARTS)-based quantitative proteomics approach, we revealed the M2 isoform of a pyruvate kinase, PKM2, as a direct binding target of butyrate. Butyrate activates PKM2 via promoting its dephosphorylation and tetramerization and thereby reprograms the metabolism of colorectal cancer cells, inhibiting the Warburg effect while favoring energetic metabolism. Our study thus provides a mechanistic link between PKM2-induced metabolic remodeling and the antitumorigenic function of butyrate and demonstrates a widely applicable approach to uncovering unknown protein targets for small molecules with biological functions.