[生酮饮食对BALB/c-nu小鼠模型神经母细胞瘤的抑制作用]
[Inhibitory effect of ketogenic diet on neuroblastoma in BALB/c-nu mouse models].
作者信息
He Jiaojiao, Lü Linya, Peng Junwei, Li Changchun, Kong Xiangru, Zhang Jun, Peng Liang
机构信息
Department of Oncology Surgery, Chongqing 400014, China.
Ministry of Education Key Laboratory of Child Development and Disorders//National Clinical Research Center for Child Health and Disorders//China International Science and Technology Cooperation Base of Child Development and Critical Disorders//Children's Hospital of Chongqing Medical University//Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Aug 30;40(8):1155-1164. doi: 10.12122/j.issn.1673-4254.2020.08.13.
OBJECTIVE
To investigate the inhibitory effect of ketogenic diet (KD) on growth of neuroblastoma in mice.
METHODS
BALB/c-nu mouse models bearing neuroblastoma xenografts were established by subcutaneous injection of human neuroblastoma cell line (SH-SY5Y). When the tumor volume reached 250 mm3, the mice were randomized into SD group with standard diet and PBS treatment, KD group with ketogenic diet and PBS treatment, and CP+KD group with ketogenic diet and cyclophosphamide (60 mg·kg·day) treatment, =8. The tumor volume, body weight, blood glucose, ketone body (β-Hydroxybutyrate) levels, and hepatic steatosis in the mice were assessed. The expressions of caspase-3 and caspase-8 were detected by Western blotting, and Ki67 expresison was detected using immunohistochemistry (IHC). Transmission electron microscopy (TEM) was employed for the autophagosomes, and the autophagic protein Beclin1, LC3A/B and P62 were detected by IHC and Western blotting.
RESULTS
On day 28 post tumor cell injection, the mice in KD and CP+KD groups could prolong the overall survival rates than that in SD group ( < 0.001). On day 22 post the injection, the tumor volume in KD group was smaller than that in SD group ( < 0.05); on 16, 19, and 22 day post the injection, the tumor volume in CP+KD group was smaller than that in SD group ( < 0.01). The mice in SD group showed greater body weight on day 19 and higher blood glucose level on day 13 post the injection than those in the other two groups ( < 0.05). Blood ketone level and hepatic steatosis score were higher and glucose ketone index (GKI) was lower in KD and CP+KD groups than those in SD group (all < 0.05). The expressions of Ki67 and apoptotic proteins were detected in the tumor tissues of all groups. TEM revealed more autophagosomes in the tumor tissues of KD group than that of SD group. P62 expression was lowered ( < 0.01) and Beclin1 and LC3A/B expressions were up-regulated in the tumor tissues of KD group ( < 0.05), which is consisitent with IHC.
CONCLUSIONS
KD has a strong anti-tumor effect in the xenograft mouse model possibly by regulating cell autophagy.
目的
探讨生酮饮食（KD）对小鼠神经母细胞瘤生长的抑制作用。
方法
通过皮下注射人神经母细胞瘤细胞系（SH-SY5Y）建立荷神经母细胞瘤异种移植瘤的BALB/c-nu小鼠模型。当肿瘤体积达到250立方毫米时，将小鼠随机分为标准饮食并给予PBS治疗的SD组、生酮饮食并给予PBS治疗的KD组和生酮饮食并给予环磷酰胺（60毫克·千克·天）治疗的CP+KD组，每组n = 8。评估小鼠的肿瘤体积、体重、血糖、酮体（β-羟基丁酸）水平和肝脂肪变性情况。采用蛋白质免疫印迹法检测半胱天冬酶-3和半胱天冬酶-8的表达，采用免疫组织化学（IHC）法检测Ki67表达。采用透射电子显微镜（TEM）观察自噬体，并通过免疫组织化学和蛋白质免疫印迹法检测自噬蛋白Beclin1、LC3A/B和P62。
结果
在肿瘤细胞注射后第28天，KD组和CP+KD组小鼠的总生存率高于SD组（P < 0.001）。在注射后第22天，KD组的肿瘤体积小于SD组（P < 0.05）；在注射后第16、19和22天，CP+KD组的肿瘤体积小于SD组（P < 0.01）。注射后第19天，SD组小鼠的体重高于其他两组（P < 0.05）；注射后第13天，SD组小鼠的血糖水平高于其他两组（P < 0.05）。KD组和CP+KD组的血酮水平和肝脂肪变性评分高于SD组，葡萄糖酮指数（GKI）低于SD组（均P < 0.05）。检测了所有组肿瘤组织中Ki67和凋亡蛋白的表达。透射电子显微镜显示，KD组肿瘤组织中的自噬体多于SD组。KD组肿瘤组织中P62表达降低（P < 0.01），Beclin1和LC3A/B表达上调（P < 0.05），这与免疫组织化学结果一致。
结论
KD在异种移植小鼠模型中具有较强的抗肿瘤作用，可能是通过调节细胞自噬实现的。
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