Ketogenic diet combined with antioxidant N-acetylcysteine inhibits tumor growth in a mouse model of anaplastic thyroid cancer
Ketogenic diet combined with antioxidant N-acetylcysteine inhibits tumor growth in a mouse model of anaplastic thyroid cancer
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DOI：
10.1016/j.surg.2019.06.042
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发表时间：
2020-01-01
期刊：
影响因子：
3.8
通讯作者：
Nehs, Matthew A.
中科院分区：
文献类型：
--
作者：
Aggarwal, Abha;Yuan, Zuliang;Nehs, Matthew A.
背景：间变性甲状腺癌是一种侵袭性、致死性恶性肿瘤。许多晚期癌症的特点是葡萄糖依赖，导致氧化应激和细胞增殖。因此，我们试图确定低糖环境（体外）或生酮饮食（体内）在与抗氧化剂n -乙酰半胱氨酸联合使用时是否能抑制间变性甲状腺癌肿瘤的生长。方法：裸鼠体内注射间变性甲状腺癌细胞8505C （n = 6/组）。1组饲喂标准日粮；2组饲喂生酮饮食；3组饲喂n -乙酰半胱氨酸标准日粮（饮水中n -乙酰半胱氨酸40 mM）；第4组饲喂含n -乙酰半胱氨酸的生酮饲粮。测量肿瘤体积、酮类和葡萄糖。对肿瘤进行H&E染色、Ki-67和Caspase 3免疫组化。8505C细胞在体外高糖（25 mM）、低糖（3 mM）、高糖+ n -乙酰半胱氨酸（200 uM）、低糖+ n -乙酰半胱氨酸培养96小时。我们进行了CyQUANT增殖（Thermo Fisher Scientific, Waltham， MA），海马糖酵解应激（Agilent, Santa Clara， CA）和反应性氧化应激试验。结果：生酮饮食加n -乙酰半胱氨酸比标准饮食(22.5 +/- 12.4 mm（3) vs 147 +/- 54.4 mm(3), P < 0.05）和标准饮食加n -乙酰半胱氨酸（P < 0.05）减少体内肿瘤体积。生酮饮食组小鼠的血酮水平显著高于标准饮食组（第5周时为1.74 mmol/L vs 0.38 mmol/L, P < 0.001）。然而，生酮饮食组和标准饮食组的血糖水平没有显著差异。与高糖相比，低糖加n -乙酰半胱氨酸培养的细胞增殖显著降低（98.1 +/- 5.0相对荧光单位vs 157.8 +/- 2.1相对荧光单位，P < 0.001）。与高糖（39.0 +/- 2.2 mpH/min/细胞vs 89.1 +/- 13.2 mpH/min/细胞，P < 0.001）和高糖加n -乙酰半胱氨酸（37.4 +/- 2.5 mpH/min/细胞vs 70.3 +/- 3.3 mpH/min/细胞，P < 0.001）相比，低糖加n -乙酰半胱氨酸降低了糖酵解功能。与高葡萄糖相比，低葡萄糖加n -乙酰半胱氨酸降低了反应性氧化应激（119 +/- 34.7相对荧光单位vs 277 +/- 16.0相对荧光单位，P = 0.014）。结论：生酮饮食或限糖饮食结合抗氧化剂n -乙酰半胱氨酸可显著降低体内和体外肿瘤生长。这些代谢疗法在间变性甲状腺癌中的应用有待进一步的研究。(C) 2019 Elsevier Inc.版权所有。
Background: Anaplastic thyroid cancer is an aggressive and fatal malignancy. Many advanced cancers are characterized by glucose dependency, leading to oxidative stress and cellular proliferation. Therefore, we sought to determine if a low glucose environment (in vitro) or a ketogenic diet (in vivo) could inhibit anaplastic thyroid cancer tumor growth when combined with the antioxidant N-acetylcysteine.Methods: In vivo, nude mice were injected with the anaplastic thyroid cancer cell line 8505C (n = 6/group). Group 1 was fed a standard diet; Group 2 was fed a ketogenic diet; Group 3 was given standard diet with N-acetylcysteine (40 mM in the drinking water); and Group 4 was fed ketogenic diet with N-acetylcysteine. Tumor volumes, ketones, and glucose were measured. H&E stains and immunohistochemistry for Ki-67 and Caspase 3 were performed on the tumors. In vitro, 8505C cells were cultured in high glucose (25 mM), low glucose (3 mM), high glucose plus N-acetylcysteine (200 uM), or low glucose plus N-acetylcysteine for 96 hours. We performed CyQUANT proliferation (Thermo Fisher Scientific, Waltham, MA), Seahorse glycolytic stress (Agilent, Santa Clara, CA), and reactive oxidative stress assays.Results: Ketogenic diet plus N-acetylcysteine decreased in vivo tumor volume compared to standard diet (22.5 +/- 12.4 mm(3) vs 147 +/- 54.4 mm(3), P < .05) and standard diet plus N-acetylcysteine (P < .05). Blood ketone levels were significantly higher for the mice in the ketogenic diet group compared to standard diet (1.74 mmol/L vs 0.38 mmol/L at week 5, P < .001). However, blood glucose levels were not significantly different between ketogenic diet and standard diet groups. Cells cultured in low glucose plus N-acetylcysteine had significantly reduced proliferation compared to high glucose (98.1 +/- 5.0 relative fluorescence units vs 157.8 +/- 2.1 relative fluorescence units, P < .001). Addition of N-acetylcysteine to low glucose lowered glycolysis function compared to high glucose (39.0 +/- 2.2 mpH/min/cell vs 89.1 +/- 13.2 mpH/min/cell, P < .001) and high glucose plus N-acetylcysteine (37.4 +/- 2.5 mpH/min/cell vs 70.3 +/- 3.3 mpH/min/cell, P < .001). Low glucose plus N-acetylcysteine decreased reactive oxidative stress compared to high glucose (119 +/- 34.7 relative fluorescence units vs 277 +/- 16.0 relative fluorescence units, P = .014).Conclusion: The combination of a ketogenic diet or glucose restriction with the antioxidant-N-acetylcysteine significantly reduced tumor growth in vivo and in vitro. Further studies are warranted to explore these metabolic therapies in anaplastic thyroid cancer. (C) 2019 Elsevier Inc. All rights reserved.