Romualdo Guilherme Ribeiro, Grassi Tony Fernando, Goto Renata Leme, Tablas Mariana Baptista, Bidinotto Lucas Tadeu, Fernandes Ana Angélica Henrique, Cogliati Bruno, Barbisan Luís Fernando
Department of Pathology, Botucatu Medical School, São Paulo State University (UNESP), Botucatu - SP, Brazil.
Department of Morphology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu - SP, Brazil.
Toxicol Lett. 2017 Nov 5;281:84-94. doi: 10.1016/j.toxlet.2017.09.015. Epub 2017 Sep 21.
This study aimed the integrative characterization of morphological, biochemical and molecular features of chemically-induced cirrhosis-associated hepatocarcinogenesis. Thus, male Wistar rats were submitted to a diethylnitrosamine (DEN)/thioacetamide (TAA)-induced model. Liver tissue was processed for global gene expression, histopathological and collagen evaluations; as well as immunohistochemical and oxidative stress analysis. Gene Ontology and functional analysis showed the upregulation of extracellular matrix deposition genes, such as collagen type I alpha 1 and 2 (Col1α1 and Col1α2) and tissue inhibitor of metalloproteinase 1 and 2 genes (Timp1 and Timp2). In agreement these findings, animals presented extensive liver cirrhosis with increased collagen deposition (Sirius red). Besides, the animals developed many glutathione S-transferase pi (GST-P)-positive preneoplastic lesions showing high cell proliferation (Ki-67), in keeping with the Gstp1 and Gstp2 increased gene expression. DEN/TAA-treated rats also showed the upregulation of tumorigenesis-related annexin A2 gene (Anxa2) and few neoplastic lesions (hepatocellular adenomas, carcinomas, and cholangiocarcinoma). In contrast, gene expression and activity of antioxidant enzymes were decreased (glutathione peroxidase, total glutathione-S-transferase, and catalase). The model featured remarkable similarities to human hepatocarcinogenesis. Our findings could bring up new molecular insights into cirrhosis-associated hepatocarcinogenesis, and provide a suitable animal model for the establishment of further diagnostic, preventive and therapeutic approaches.
本研究旨在对化学诱导的肝硬化相关肝癌发生的形态学、生化和分子特征进行综合表征。因此，将雄性Wistar大鼠用于二乙基亚硝胺（DEN）/硫代乙酰胺（TAA）诱导的模型。对肝组织进行整体基因表达、组织病理学和胶原评估，以及免疫组织化学和氧化应激分析。基因本体论和功能分析显示细胞外基质沉积基因上调，如I型胶原α1和α2（Col1α1和Col1α2）以及金属蛋白酶组织抑制剂1和2基因（Timp1和Timp2）。与这些发现一致，动物出现广泛的肝硬化，胶原沉积增加（天狼星红染色）。此外，动物出现许多谷胱甘肽S-转移酶pi（GST-P）阳性的癌前病变，显示细胞增殖高（Ki-67），这与Gstp1和Gstp2基因表达增加一致。DEN/TAA处理的大鼠还显示肿瘤发生相关膜联蛋白A2基因（Anxa2）上调，并有少量肿瘤性病变（肝细胞腺瘤、癌和胆管癌）。相比之下，抗氧化酶的基因表达和活性降低（谷胱甘肽过氧化物酶、总谷胱甘肽-S-转移酶和过氧化氢酶）。该模型与人类肝癌发生有显著相似之处。我们的发现可为肝硬化相关肝癌发生带来新的分子见解，并为建立进一步的诊断、预防和治疗方法提供合适的动物模型。
