化学致癌物二乙基亚硝胺（DEN）经常被用来诱发小鼠肝癌。奇怪的是，一些实验室报告称，从肝细胞中去除癌蛋白会加剧 DEN 诱发的 HCC，其机制尚不清楚。本研究旨在破译癌蛋白肿瘤抑制作用背后的分子机制。我们培育了肝细胞特异性删除 Met、Ptpn11/Shp2、Ikkβ 或 Ctnnb1/β-catenin 的突变小鼠系，并评估了 DEN 在野生环境中诱导的肿瘤发生。型和突变型小鼠。为了系统地检查遗传和分子信号传导的改变，我们对癌前阶段和既定癌症阶段收集的肝脏样本进行了全外显子组和 RNA 测序。尽管野生型和突变型小鼠中 DEN 诱导的肿瘤的突变谱几乎没有差异，但癌蛋白消除增加了 DEN 诱导的突变负担，特别是在 Shp2 缺陷的肿瘤中。 RNA测序揭示了信号通路的多种变化，特别是上皮-间质转化、细胞迁移和肿瘤转移的上调，以及受癌蛋白消融影响的小分子代谢的下调。我们确定了与肝脏先天免疫相关并与肝脏肿瘤发生有关的关键分子和途径。此外，我们还发现人类 HCC 数据库中一些 miRNA 的表达发生显着变化。在癌蛋白消融中观察到的 DEN 诱导的 HCC 进展加剧可能是由常见且不同的基因组和信号传导改变引起的。这项研究揭示了肝癌发生的新复杂性，并阐明了肿瘤演变和复发的分子机制。版权所有 © 2024 美国肝病研究协会。

The chemical carcinogen diethylnitrosamine (DEN) is often used to induce HCC in mice. Curiously, several labs have reported that the removal of oncoproteins from hepatocytes exacerbated DEN-induced HCC, with mechanisms unknown. This study aimed at deciphering molecular mechanisms underlying the tumor suppressive effect of oncoproteins.We generated mutant mouse lines with hepatocyte-specific deletions of Met, Ptpn11/Shp2, Ikkβ, or Ctnnb1/β-catenin and assessed DEN-induced tumorigenesis in the wild-type and mutant mice. To systematically examine genetic and molecular signaling alterations, we performed whole exome and RNA-sequencing on liver samples collected at the pre-cancer and established cancer stages. Although the mutational profiles of DEN-induced tumors were barely different in wild-type and mutant mice, oncoprotein ablation increased DEN-induced mutational burdens, especially in Shp2-deficient tumors. RNA-sequencing revealed multiple changes in signaling pathways, in particular, upregulated epithelial-mesenchymal transition, cell migration, and tumor metastasis, as well as downregulated small molecule metabolism that was affected by oncoprotein ablation. We identified key molecules and pathways that are associated with hepatic innate immunity and implicated in liver tumorigenesis. In addition, we unveiled markedly changed expression of a few miRNAs in the human HCC database.The aggravation of DEN-induced HCC progression seen on oncoprotein ablation could be caused by common and distinct genomic and signaling alterations. This study reveals a new level of complexity in hepatocarcinogenesis and elucidates molecular mechanisms underlying tumor evolution and recurrence.Copyright © 2024 American Association for the Study of Liver Diseases.