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International Journal of Molecular... May 2023This study aimed to analyze the biochemical, histological, and gene expression alterations produced in a hepatocarcinogenesis model induced by the chronic administration...
This study aimed to analyze the biochemical, histological, and gene expression alterations produced in a hepatocarcinogenesis model induced by the chronic administration of diethylnitrosamine (DEN) and 2-acetylaminofluorene (2-AAF) in Wistar rats. Thirteen rats weighing 180 to 200 g were divided into two groups: control and treated. Rats in the treated group were administered an intraperitoneal (i.p.) injection of DEN (50 mg/kg/week) and an intragastric (i.g.) dose of 2-AAF (25 mg/kg/week) for 18 weeks. The treated group had significant increases in their total cholesterol, HDL-C, AST, ALT, ALKP, and GGT levels. Furthermore, a histological analysis showed the loss of normal liver architecture with nuclear pleomorphism in the hepatocytes, atypical mitosis, and fibrous septa that were distributed between the portal triads and collagen fibers through the hepatic sinusoids. The gene expressions of 24 genes related to fibrosis, inflammation, apoptosis, cell growth, angiogenesis, lipid metabolism, and alpha-fetoprotein () were analyzed; only , , , , , , and showed significant differences when both groups were compared. Additionally, lung histopathological alterations were found in the treated group, suggesting metastasis. In this model, the chronic administration of DEN+2-AAF induces characteristic alterations of hepatocellular carcinoma in Wistar rats without gene expression changes, highlighting different signatures in hepatocellular carcinoma heterogeneity.
Topics: Rats; Animals; Carcinoma, Hepatocellular; Rats, Wistar; Liver; 2-Acetylaminofluorene; Diethylnitrosamine; alpha-Fetoproteins; Liver Neoplasms; Liver Neoplasms, Experimental
PubMed: 37176094
DOI: 10.3390/ijms24098387 -
Experimental and Molecular Pathology Dec 2021This study was intended (1) to develop a robust animal model for hepatocellular carcinoma (HCC) research, in which HCC tumors develop in a background of fibrosis or...
This study was intended (1) to develop a robust animal model for hepatocellular carcinoma (HCC) research, in which HCC tumors develop in a background of fibrosis or cirrhosis; and (2) to explore time-dependent regulatory changes in key molecular markers during disease advancement and HCC development. With the aim of establishing such HCC model, male Sprague-Dawley rats were injected with diethylnitrosamine (DEN) at a dose of 30 mg/kg twice a week for 10 weeks then once a week from 12th to 16th weeks. The rats were kept under observation until 18th week. At defined time intervals (2nd, 4th, 12th, and 18th week), serum biomarkers and microscopic components of tissue samples were used to investigate the chronic progression of liver disease, while gene and protein analysis was used to monitor expression patterns during HCC development. DEN-intoxicated rats manifested inflammation at week 4, fibrosis at week 12 and cirrhosis with early HCC tumors at week 18. Molecular analysis revealed that key markers of inflammation (Il-1β, Il-6, and Tnf-α), fibrosis (Tgf-β1, Col1α1, Col3α1, and Timp-1), and angiogenesis (Hif1-α and Vegf) were promptly (P ≤ 0.001) up-regulated at week 4, week 12 and week 18, respectively. Oxidative stress (iNos, Cyp2e1, and Sod1) and pro-apoptotic (Bax) markers showed significant upregulation from week 4 to week 12. However, Sod1 and Bax expressions dropped after week 12 and reached a minimum at 18th week. Strikingly, expressions of anti-apoptotic (Bcl-2) and cell proliferation (Pcna, Hgf, and Afp) markers were abruptly increased at week 18. Collectively, we describe an 18-week HCC model in DEN-intoxicated rats that exhibit chronic inflammation, oxidative imbalance, advance fibrosis/cirrhosis, halted apoptosis, and angiogenic sprouting, progressively.
Topics: Animals; Apoptosis; Carcinogenesis; Carcinoma, Hepatocellular; Cell Proliferation; Diethylnitrosamine; Disease Models, Animal; Fibrosis; Gene Expression Regulation, Neoplastic; Humans; Inflammation; Liver; Liver Cirrhosis; Liver Neoplasms; Neoplasm Proteins; Neovascularization, Pathologic; Rats
PubMed: 34699901
DOI: 10.1016/j.yexmp.2021.104715 -
Food and Chemical Toxicology : An... Feb 2019The liver is the most essential organ of the body performing vital functions. Hepatic disorders affect the physiological and biochemical functions of the body. These... (Review)
Review
The liver is the most essential organ of the body performing vital functions. Hepatic disorders affect the physiological and biochemical functions of the body. These disorders include hepatitis B, hepatitis C, alcoholic liver disease (ALD), nonalcoholic fatty liver disease (NAFLD), liver cirrhosis, hepatic failure and hepatocellular carcinoma (HCC). Drugs related hepatotoxicity is one of the major challenges facing by clinicians as it is a leading cause of liver failure. During post-marketing surveillance studies, detection and reporting of drug-induced hepatotoxicity may lead to drug withdrawal or warnings. Several mechanisms are involved in hepatotoxicity such as cell membrane disruption, initiating an immune response, alteration of cellular pathways of drug metabolism, accumulation of reactive oxygen species (ROS), lipid peroxidation and cell death. Curcumin, the active ingredient of turmeric and exhibits therapeutic potential for the treatment of diabetes, cardiovascular disorders and various types of cancers. Curcumin is strong anti-oxidant and anti-inflammatory effects and thus it possesses hepatoprotective properties. Despite its low bioavailability, its hepatoprotective effects have been studied in various protocols of hepatotoxicity including acetaminophen, alcohol, lindane, carbon tetrachloride (CCL), diethylnitrosamine and heavy metals induced hepatotoxicities. This report reviews the hepatoprotective effects of curcumin with a focus on its mechanistic insights in various hepatotoxic protocols.
Topics: Acetaminophen; Animals; Anti-Inflammatory Agents; Antioxidants; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Curcumin; Diethylnitrosamine; Hexachlorocyclohexane; Humans; Liver; Metals, Heavy; Oxidative Stress; Protective Agents
PubMed: 30529260
DOI: 10.1016/j.fct.2018.12.002 -
Journal of Basic and Clinical... Jun 2021For designing early treatment for liver cancer, it is important to prepare an animal model to evaluate cancer prevention treatment by using inflammation disease. The...
OBJECTIVES
For designing early treatment for liver cancer, it is important to prepare an animal model to evaluate cancer prevention treatment by using inflammation disease. The hepatocarcinogenic N-Nitrosodiethylamine (NDEA) has been reportedly able to produce free radicals that cause liver inflammation leading to liver carcinoma. This study aimed to evaluate the inflammation disease model of mice induced with hepatocarcinogenic NDEA for five weeks induction.
METHODS
The BALB-c mice were induced with NDEA 25 mg/kg of body weight once a week for five weeks intraperitonially and it was then evaluated for the body weight during study periods. The mice were then sacrificed and excised for evaluating their organs including physical and morphological appearances and histopathology evaluations.
RESULTS
The results showed a significant decrease of body weight of mice after five times induction of 25 mg NDEA/kgBW per week intraperitonially. Different morphological appearances and weight of mice organs specifically for liver and spleen had also been observed. The histopathology examination showed that there were hepatic lipidosis and steatohepatitis observed in liver and spleen, respectively that might indicate the hepatocellular injury.
CONCLUSIONS
It can be concluded that inducing mice with NDEA intraperitonially resulted in fatty liver disease leading to progress of cancer disease.
Topics: Animals; Body Weight; Diethylnitrosamine; Inflammation; Liver; Liver Neoplasms, Experimental; Mice; Mice, Inbred BALB C
PubMed: 34214328
DOI: 10.1515/jbcpp-2020-0475 -
Carcinogenesis Oct 2015MiR-17-92 cluster is an oncogenic miRNA cluster that is implicated in several cancers, although its role in hepatocarcinogenesis has not been clearly defined. In this...
MiR-17-92 cluster is an oncogenic miRNA cluster that is implicated in several cancers, although its role in hepatocarcinogenesis has not been clearly defined. In this study, we show that the miR-17-92 cluster is highly expressed in human hepatocellular carcinoma (HCC) tissues compared to the non-tumorous liver tissues by RT-PCR and in situ hybridization analyses. Increased miR-17-92 cluster expression in HCC tissues was further confirmed by analysis of the RNA-sequencing data of 319 patients available from the Cancer Genome Atlas (TCGA) Data Portal (https://tcga-data.nci.nih.gov/tcga/). To create an animal model that resembles enhanced miR-17-92 in the liver, we developed liver-specific miR-17-92 transgenic mice and the animals were treated with the hepatic carcinogen, diethylnitrosamine (DEN). We observed that the liver-specific miR-17-92 transgenic mice showed significantly increased hepatocellular cancer development compared to the matched wild-type control mice. Forced overexpression of the miR-17-92 cluster in cultured human hepatocellular cancer cells enhanced tumor cell proliferation, colony formation and invasiveness in vitro, whereas inhibition of the miR-17-92 cluster reduced tumor cell growth. By analyzing the miRNA and mRNA sequencing data from the 312 hepatocellular cancer patients available from the TCGA database, we observed that the expression levels of the miR-17-92 cluster members and host gene in the tumor tissues are negatively correlated with several target genes, including CREBL2, PRRG1, NTN4. Our findings demonstrate an important role of the miR-17-92 cluster in hepatocarcinogenesis and suggest the possibility of targeting this pivotal miRNA cluster for potential therapy.
Topics: Animals; Carcinogenesis; Carcinoma, Hepatocellular; Cell Proliferation; Diethylnitrosamine; Disease Models, Animal; Gene Expression Regulation, Neoplastic; Hep G2 Cells; High-Throughput Nucleotide Sequencing; Humans; Liver Neoplasms; Mice; Mice, Transgenic; MicroRNAs; RNA, Long Noncoding
PubMed: 26233958
DOI: 10.1093/carcin/bgv112 -
International Journal For Vitamin and... Oct 2022β-Carotene exhibits antioxidant and hepatoprotective activities a multitude of biochemical mechanisms. However, the action mechanism involved in antioxidant and...
β-Carotene exhibits antioxidant and hepatoprotective activities a multitude of biochemical mechanisms. However, the action mechanism involved in antioxidant and anti-inflammatory effects of this carotene in chronic liver diseases is not fully understood. In the present investigation, we have attempted to outline a plausible mechanism of β-carotene action against liver fibrosis in albino Wistar rats. To induce hepatic fibrosis, diethylnitrosamine (DEN) was administered in experimental rats for two weeks. DEN treated rats were divided into four groups, wherein each group comprised of five rats. β-Carotene supplement attenuated DEN-induced elevation in LFT markers (P < 0.05); averted depletion of glycogen (24%, P < 0.05) and, increased nitrite (P < 0.05), hydroxyproline (~67%, P < 0.05) and collagen levels (~65%, P < 0.05). Confocal microscopy of tissue sections stained with picrosirius red revealed accrued collagen in DEN-administered group, which was found to be reduced by β-carotene supplementation. Furthermore, β-carotene decreased the expression of iNOS/NOS-2 and NF-κB, as revealed by immunohistochemistry and Western immunoblotting. Collectively, these results demonstrate that β-carotene mitigates experimental liver fibrosis inhibition of iNOS and NF-κB . Thus, β-carotene may be suggested as a possible nutraceutical to curb experimental liver fibrosis.
Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Diethylnitrosamine; Glycogen; Hydroxyproline; Inflammation; Liver; Liver Cirrhosis; NF-kappa B; Nitrites; Rats; Rats, Wistar; beta Carotene
PubMed: 32686990
DOI: 10.1024/0300-9831/a000665 -
Journal of Translational Medicine Jan 2024Hepatocellular carcinoma (HCC) remains a leading life-threatening health challenge worldwide, with pressing needs for novel therapeutic strategies. Sphingosine kinase 1...
BACKGROUND
Hepatocellular carcinoma (HCC) remains a leading life-threatening health challenge worldwide, with pressing needs for novel therapeutic strategies. Sphingosine kinase 1 (SphK1), a well-established pro-cancer enzyme, is aberrantly overexpressed in a multitude of malignancies, including HCC. Our previous research has shown that genetic ablation of Sphk1 mitigates HCC progression in mice. Therefore, the development of PF-543, a highly selective SphK1 inhibitor, opens a new avenue for HCC treatment. However, the anti-cancer efficacy of PF-543 has not yet been investigated in primary cancer models in vivo, thereby limiting its further translation.
METHODS
Building upon the identification of the active form of SphK1 as a viable therapeutic target in human HCC specimens, we assessed the capacity of PF-543 in suppressing tumor progression using a diethylnitrosamine-induced mouse model of primary HCC. We further delineated its underlying mechanisms in both HCC and endothelial cells. Key findings were validated in Sphk1 knockout mice and lentiviral-mediated SphK1 knockdown cells.
RESULTS
SphK1 activity was found to be elevated in human HCC tissues. Administration of PF-543 effectively abrogated hepatic SphK1 activity and significantly suppressed HCC progression in diethylnitrosamine-treated mice. The primary mechanism of action was through the inhibition of tumor neovascularization, as PF-543 disrupted endothelial cell angiogenesis even in a pro-angiogenic milieu. Mechanistically, PF-543 induced proteasomal degradation of the critical glycolytic enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3, thus restricting the energy supply essential for tumor angiogenesis. These effects of PF-543 could be reversed upon S1P supplementation in an S1P receptor-dependent manner.
CONCLUSIONS
This study provides the first in vivo evidence supporting the potential of PF-543 as an effective anti-HCC agent. It also uncovers previously undescribed links between the pro-cancer, pro-angiogenic and pro-glycolytic roles of the SphK1/S1P/S1P receptor axis. Importantly, unlike conventional anti-HCC drugs that target individual pro-angiogenic drivers, PF-543 impairs the PFKFB3-dictated glycolytic energy engine that fuels tumor angiogenesis, representing a novel and potentially safer therapeutic strategy for HCC.
Topics: Animals; Humans; Mice; Angiogenesis; Carcinoma, Hepatocellular; Diethylnitrosamine; Endothelial Cells; Liver Neoplasms; Methanol; Neovascularization, Pathologic; Phosphofructokinase-2; Phosphotransferases (Alcohol Group Acceptor); Pyrrolidines; Sphingosine-1-Phosphate Receptors; Sulfones
PubMed: 38200582
DOI: 10.1186/s12967-023-04830-z -
Cells Sep 2022MiR-22 is mostly considered as a hepatic tumor-suppressor microRNA based on in vitro analyses. Yet, whether miR-22 exerts a tumor-suppressive function in the liver has...
MiR-22 is mostly considered as a hepatic tumor-suppressor microRNA based on in vitro analyses. Yet, whether miR-22 exerts a tumor-suppressive function in the liver has not been investigated in vivo. Herein, in silico analyses of miR-22 expression were performed in hepatocellular carcinomas from human patient cohorts and different mouse models. Diethylnitrosamine-induced hepatocellular carcinomas were then investigated in lean and diet-induced obese miR-22-deficient mice. The proteome of liver tissues from miR-22-deficient mice prior to hepatocellular carcinoma development was further analyzed to uncover miR-22 regulated factors that impact hepatocarcinogenesis with miR-22 deficiency. MiR-22 downregulation was consistently observed in hepatocellular carcinomas from all human cohorts and mouse models investigated. The time of appearance of the first tumors was decreased and the number of tumoral foci induced by diethylnitrosamine was significantly increased by miR-22-deficiency in vivo, two features which were further drastically exacerbated with diet-induced obesity. At the molecular level, we provide evidence that the loss of miR-22 significantly affects the energetic metabolism and mitochondrial functions of hepatocytes, and the expression of tumor-promoting factors such as thrombospondin-1. Our study demonstrates that miR-22 acts as a hepatic tumor suppressor in vivo by restraining pro-carcinogenic metabolic deregulations through pleiotropic mechanisms and the overexpression of relevant oncogenes.
Topics: Animals; Carcinogenesis; Carcinoma, Hepatocellular; Diethylnitrosamine; Disease Models, Animal; Fatty Liver; Humans; Liver Neoplasms; Mice; MicroRNAs; Proteome; Thrombospondins
PubMed: 36139435
DOI: 10.3390/cells11182860 -
Analytical Chemistry May 2020Diversified oxidized-lipid molecules are responsible for inflammation and cell death, including ferroptosis. Lipid radicals are the source of these oxidized lipids,...
Diversified oxidized-lipid molecules are responsible for inflammation and cell death, including ferroptosis. Lipid radicals are the source of these oxidized lipids, which are the initial key molecules in the lipid peroxidation chain reaction. However, owing to their extremely high reactivity and short half-life, an established detection technique is not available. Here, we propose a high-performance liquid chromatography fluorometry and high-resolution tandem mass spectrometry system combined with a fluorescent probe as a structural analysis method for lipid-derived radicals. We detected 132 lipid-derived radicals, including 111 new species, from five polyunsaturated fatty acids. In addition, a database was constructed for which the initial fatty acid could be determined using the radical structure. Further, 12 endogenous lipid-derived radicals were identified in carcinogen-induced liver cancer mouse models. Therefore, this method and its corresponding database will provide novel insights into mechanisms underlying the lipid peroxidation, including the associated inflammation and ferroptosis.
Topics: Animals; Diethylnitrosamine; Disease Models, Animal; Free Radicals; Injections, Intraperitoneal; Lipids; Liver Neoplasms; Male; Mice; Mice, Inbred C57BL; Molecular Structure
PubMed: 32311262
DOI: 10.1021/acs.analchem.0c00053 -
Toxicology Oct 2022This study aims to evaluate in vivo protective effects of eumelanin (EU) on diethylnitrosamine (DEN)-induced liver injury. Wistar albino male rats were divided into 6...
This study aims to evaluate in vivo protective effects of eumelanin (EU) on diethylnitrosamine (DEN)-induced liver injury. Wistar albino male rats were divided into 6 groups (n = 6), Control, DMSO, DEN, DEN + EU10, DEN + EU15, and DEN + EU20. Animals in the DEN group were injected i.p a single dose of 200 mg/kg DEN, DEN + EU10 group was given 10 mg/kg EU, DEN + EU15 group was given 15 mg/kg, DEN + EU20 group was given 20 mg/kg EU for a week. The results showed that there was no significant difference in vessel volume density between the groups. Inflammatory cell infiltration, hydropic degeneration, and necrotic cells were observed in the DEN group, and these histopathological changes were significantly reduced in all treatment groups. Although there was a low intensity of PAS-positive staining in the DEN groups, moderate staining was observed in the treatment groups. While Caspase-3, PCNA, TNF-α, and IL-6 expressions increased in the DEN group, their expressions decreased in the EU-treated groups. DEN increased AST, ALT, and MDA levels and decreased CAT levels. In particular, the EU10 dose significantly improved these parameters. The present study revealed that eumelanin has protective effects against DEN-induced liver injury.
Topics: Animals; Caspase 3; Chemical and Drug Induced Liver Injury, Chronic; Diethylnitrosamine; Dimethyl Sulfoxide; Interleukin-6; Liver; Melanins; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha
PubMed: 36113623
DOI: 10.1016/j.tox.2022.153311