该研究旨在通过转录组测序技术筛选出驴油生酮饮食（Donkey Oil Ketogeni Diet，DOKD）影响CT26+结肠癌肿瘤生长的相关基因，挖掘对DOKD响应的基因及其代谢通路，为解析驴油生酮饮食抑制肿瘤生长的机制提供依据。该研究选取一致性较好的CT26+结肠癌BALB/c模型小鼠，随机分为2组，每组3个重复，分别饲喂驴油生酮饮食（DOKD）与正常饮食（ND）。饲喂10 d后处死摘取肿瘤组织，测量肿瘤的重量及体积，并利用RNA-Seq技术和生物信息分析方法进行转录组测序及结果分析。DOKD组与ND组小鼠体重无显著差异，肿瘤明显小于ND组。与ND组相比，DOKD组共18个基因差异表达，其中12个DEGs上调，6个DEGs下调。这些DEGs注释到170条GO条目，其中生物过程（BP）类别124条，细胞组成（CC）类别2条，分子功能（MF）类别44条；涉及18条KEGG通路，主要富集在IL-17信号通路、趋化因子信号通路、细胞因子受体相互作用信号通路、肿瘤坏死因子信号通路等重要通路。此外，该研究通过Western blot进一步研究了TLR4/NF-κB信号通路在在肿瘤中的作用。综上，DOKD对CT26+结肠癌肿瘤有明显抑制作用，该作用可能与IL-17信号通路、趋化因子信号通路、细胞因子受体相互作用信号通路、TNF信号通路等通路密切相关。

Screening of the genes related to the donkey oil ketogenic diet (DOKD) that affects the growth of CT26+ colon cancer tumors using transcriptome sequencing technology was investigated in this study. Mining the genes and metabolic pathways that respond to DOKD provides a basis for analyzing the mechanism of DOKD in inhibiting tumor growth. BALB/c model mice with CT26+ colon cancer with high consistency were selected and randomly divided into two groups with three replicates, and they were fed either a DOKD or normal diet (ND). The animals were sacrificed after 10 days of treatment. Subsequently, the tumors were excised, and tumor weights and volumes were recorded for analysis. RNA-Seq technology and bioinformatic analysis were used for transcriptome sequencing and results analysis. No significant difference was found in the body weight between the DOKD and ND groups, and the tumors were significantly smaller in the DOKD than those in the ND group. Compared with the ND group, a total of 18 genes were differentially expressed in the DOKD group, of which 12 differentially expressed genes (DEGs) were upregulated, and 6 DEGs were down regulated. These DEGs were annotated into a total of 170 GO items, including 124 in the biological process category, 44 in the molecular function category, and 2 in the cellular composition category. A total of 18 KEGG pathways were involved, mainly enriched in the interleukin (IL)-17 signaling, chemokine signaling, cytokine receptor interaction signaling, tumor necrosis factor signaling, and other important pathways. In addition, this study further examined the role of the TLR4/NF-κB signaling pathway in tumors using western blotting. In summary, DOKD has a significant inhibitory effect on CT26+ colon cancer tumors, and this effect may be closely related to the IL-17, chemokine, cytokine receptor interaction, and tumor necrosis factor signaling pathways, among others.

山东省现代农业产业技术体系驴产业创新团队项目资助（SDAIT-27）；山东省乡村振兴科技创新提振行动计划项目（2021TZXD012）；聊城大学博士科研启动项目（318052120）；聊城大学畜牧学学科开放课题（319312101-03）