DT-13 inhibited the proliferation of colorectal cancer via glycolytic metabolism and AMPK/mTOR signaling pathway
DT-13 inhibited the proliferation of colorectal cancer via glycolytic metabolism and AMPK/mTOR signaling pathway
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DT-13通过糖酵解代谢和AMPK/mTOR信号通路抑制结直肠癌增殖
DOI：
10.1016/j.phymed.2018.09.003
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发表时间：
2019-02-15
期刊：
影响因子：
7.9
通讯作者：
Sun, Li
中科院分区：
文献类型：
--
作者：
Wei, Xiaohui;Mao, Tingting;Sun, Li
背景资料：肿瘤的标志是细胞代谢的重新编程，糖酵解代谢增加是人类恶性肿瘤的生理特征。麦冬皂苷单体13（Saponin monomer 13 of the dwarf Lilyturf tuber，DT-13）是麦冬中主要的甾体皂苷类化合物，具有抗炎、抗肿瘤等作用，但对正常组织的毒性较低。目的：本研究旨在探讨DT-13对大肠癌细胞葡萄糖代谢的影响，并探讨其是否参与了DT-13的抗肿瘤作用。方法：采用集落形成实验检测DT-13在2.5、5、10 μ M浓度下对大肠癌细胞的增殖抑制作用。Annexin V/PI染色检测细胞凋亡，PI染色检测细胞周期阻滞。糖酵解代谢的遗传抑制通过敲低GLUT 1来进行。采用小鼠结直肠癌原位移植模型，评价DT-13（0.625、1.25、2.5 mg/kg）的体内抗肿瘤作用。采用C57 BL/6 J APC（min）小鼠模型，观察DT-13（10 mg/kg）对小鼠脑缺血再灌注损伤的预防作用。采用实时荧光定量PCR、免疫印迹和免疫组织化学方法检测糖酵解相关关键酶和AMPK通路的活性。结果：DT-13对细胞增殖有明显的抑制作用，且呈剂量依赖性。DT-13抑制葡萄糖摄取、ATP生成并减少乳酸产生。此外，DT-13在mRNA和蛋白水平上显著抑制GLUT 1表达。GLUT 1的敲低导致DT-13处理后葡萄糖摄取的抑制减少。此外，GLUT 1的缺失降低了DT-13对肿瘤生长的抑制率。原位种植结直肠癌小鼠模型进一步证实DT-13通过体内阻断GLUT 1抑制结直肠癌生长。此外，C57 BL/6 J APC（min）小鼠模型显示DT-13显著减少肠道自发性腺瘤的总数，这进一步证实了DT-13在结直肠癌中的抗肿瘤活性。结论：DT-13是一种有效的抗大肠癌药物。
Background: Emerging hallmark of cancer is reprogrammed cellular metabolism, increased glycolytic metabolism is physiological characteristic of human malignant neoplasms. Saponin monomer 13 of the dwarf lilyturf tuber (DT-13) is the main steroidal saponin from Liriopes Radix, which has been reported to exert anti-inflammation and anti-tumor activities but low toxicity to normal tissue. However, the effect of DT-13 on metabolism process is still unclear.Purpose: This study aims to characterize the role of DT-13 in glucose metabolism in colorectal cancer cells, and investigate whether the metabolism process is involved in the anti-cancer response of DT-13.Methods: Colony formation assay was employed to determine anti-proliferative effect induced by DT-13 at 2.5, 5, 10 mu M. Apoptosis and cell cycle arrest were detected by Annexin V/PI staining and PI staining, respectively. Genetic inhibition of glycolytic metabolism was carried out by knockdown of GLUT1. Orthotopic implantation mouse model of colorectal cancer was used to assess in vivo antitumor effect of DT-13 (0.625, 1.25, 2.5 mg/kg). The chemoprevention effect of DT-13 (10mg/kg) was evaluated by using C57BL/6J APC(min) mice model. Glycolytic-related key enzymes and AMPK pathway were detected by using quantitative real-time PCR, western blotting, and immunohistochemical staining.Results: Our results showed that cell proliferation was significantly inhibited by DT-13 in a dose-dependent manner. DT-13 inhibited glucose uptake, ATP generation, and reduced lactate production. Furthermore, DT-13 remarkably inhibited GLUT1 expression in both mRNA and protein levels. Knocking down of GLUT1 led to reduced inhibition of glucose uptake after DT-13 treatment. Moreover, deletion of GLUT1 decreased inhibitory ratio of DT-13 on cancer growth. Orthotopic implantation mouse model of colorectal cancer further confirmed that DT-13 inhibited colorectal cancer growth via blocking GLUT1 in vivo. In addition, C57BL/6J APC(min) mice model revealed that DT-13 dramatically reduced the total number of spontaneous adenomas in intestinal, which further confirmed the anti-tumor activity of DT-13 in colorectal cancer. Furthermore, the mechanistically investigation showed DT-13 activated AMPK and inhibited m-TOR to block cancer growth in vitro.Conclusion: DT-13 is a potent anticancer agent for colorectal cancer.