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Malignant Transformation into Liver Cancer  \nXin Yin 1,†, Yu-Sang Li 1,†, Sha-Zhou Ye 2, Ting Zhang 1, Yi-Wen Zhang 1, Yang Xi 2, * and He-Bin Tang 1, *  \nCitation: Yin, X.; Li, Y.-S.; Ye, S.-Z.; Zhang, T.; Zhang, Y.-W.; Xi, Y.; Tang, H.-B. Promotion Effect of Coexposure to a High-Fat Diet and Nano-Diethylnitrosamine on the Progression of Fatty Liver Malignant Transformation into Liver Cancer. Int.  \nJ. Mol. Sci. 2023, 24, 14162. [https://](https://)[ ](https://)[doi.org/10.3390/ijms241814162](doi.org/10.3390/ijms241814162)  \nAcademic Editor: Amedeo Lonardo  \nReceived: 25 August 2023  \nRevised: 9 September 2023  \nAccepted: 12 September 2023  \nPublished: 15 September 2023  \nCopyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) .  \n1 Lab of Hepatopharmacology and Ethnopharmacology, School of Pharmaceutical Sciences, South-Central Minzu University, No. 182, Minyuan Road, Wuhan 430074, China; [kno521521@163.com](kno521521@163.com) (X.Y.); [liys2006@mail.scuec.edu.cn](liys2006@mail.scuec.edu.cn) (Y.-S.L.); [kinima988@outlook.com](kinima988@outlook.com) (T.Z.); [2021120621@mail.scuec.edu.cn](2021120621@mail.scuec.edu.cn) (Y.-W.Z.)  \n2 Institute of Biochemistry and Molecular Biology, School of Medicine, Ningbo University, No. 818 Fenghua Road, Jiangbei District, Ningbo 315211, China; [yeshazhou@foxmail.com](yeshazhou@foxmail.com)  \n* Correspondence: [xiyang@nbu.edu.cn](xiyang@nbu.edu.cn) (Y.X.); [hbtang2006@mail.scuec.edu.cn](hbtang2006@mail.scuec.edu.cn) (H.-B.T.);  \nTel.: +86-574-8760-0754 (Y.X.); +86-27-6784-2332 (H.-B.T.)† These authors contributed equally to this work.  \nAbstract: Overconsumption of high-fat foods increases the risk of fatty liver disease (FLD) and liver cancer with long pathogenic cycles. It is also known that the intake of the chemical poison nitrosamine and its nanopreparations can promote the development of liver injuries, such as FLD, and hepatic fibrosis, and significantly shorten the formation time of the liver cancer cycle. The present work confirmed that the coexposure of a high-fat diet (HFD) and nano-diethylnitrosamine (nano-DEN) altered the tumor microenvironment and studied the effect of this coexposure on the progression of fatty liver malignant transformation into liver cancer. Gene transcriptomics and immunostaining were used to evaluate the tumor promotion effect of the coexposure in mice. After coexposure treatment, tumor nodules were obviously increased, and inflammation levels were elevated. The liver transcriptomics analysis showed that the expression levels of inflammatory, fatty, and fibrosis-related factors in the coexposed group were increased in comparison with the nano-DEN-and high-fat-alone groups. The Kyoto Encyclopedia of Genes and Genomes (KEGG) results showed that coexposure aggravated the high expression of genes related to the carcinomatous pathway and accelerated the formation of the tumor microenvironment. The immunohistochemical staining results showed that the coexposure significantly increased the abnormal changes in proteins related to inflammation, proliferation, aging, and hypoxia in mouse liver tissues. The coexposure of high fat and nano-DEN aggravated the process of steatosis and carcinogenesis. In conclusion, the habitual consumption of pickled foods containing nitrosamines in a daily HFD significantly increases the risk of liver pathology lesions progressing from FLD to liver cancer.  \nKeywords: high-fat diet; nano-diethylnitrosamine; fatty liver; liver cancer; transcriptomics  \n1. Introduction  \nLiver cancer is one of the top cancer killers worldwide, with a","cbCainVkdqRFNchc","https://ap.wps.com/l/cbCainVkdqRFNchc","pdf",4422804,"English","# Abstract\n# Introduction","[{\"question\":\"What was the main question of the study?\",\"answer\":\"How coexposure to a high-fat diet and nano-diethylnitrosamine affects progression from fatty liver malignant transformation into liver cancer.\"},{\"question\":\"Which experimental approaches were used to assess tumor promotion?\",\"answer\":\"Gene transcriptomics and immunostaining were used to evaluate the tumor promotion effect in mice.\"},{\"question\":\"What were the key findings after coexposure?\",\"answer\":\"Coexposure increased tumor nodules and inflammation, upregulated inflammatory/fatty/fibrosis-related factors, intensified carcinogenic pathway gene expression, and increased abnormal protein changes related to inflammation, proliferation, aging, and hypoxia.\"}]","Promotion Effect of Coexposure to a High-Fat Diet and Nano-Diethylnitrosamine on the Progression of Fatty Liver Malignant Transformation into Liver Cancer | PDF",1790223949]