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BioMed Research International 2017Aberrant activation of the TGF-1/Smad pathway contributes to the activation of hepatic stellate cells (HSCs). MicroRNA-195 has been shown to regulate the activation of...
BACKGROUND AND AIM
Aberrant activation of the TGF-1/Smad pathway contributes to the activation of hepatic stellate cells (HSCs). MicroRNA-195 has been shown to regulate the activation of HSCs. The aim of this study was to investigate the role of miRNA-195 in HSCs activation.
METHODS
A liver fibrotic rat model induced by diethylnitrosamine was established. Dual luciferase reporter assays were performed to verify that Smad7 was the target of miRNA-195. The expression levels of miR-195, Smad7, and -SMA in HSC-T6 transfected, respectively, with miR-195 mimic, inhibitor, or control were measured by qRT-PCR. The protein expression of Smad7 was detected by Western blot analysis.
RESULTS
Enhanced miR-195 and decreased Smad7 were observed in diethylnitrosamine-induced liver fibrotic rats ( < 0.05). Dual luciferase reporter assays showed that the miR-195 mimic significantly suppressed the luciferase activity of a reporter plasmid carrying the binding site of miR-195 on the 3'UTR of Smad7 ( < 0.05). The miR-195 mimics activated HSCs, further elevated miR-195 and -SMA ( < 0.01), and reduced the Smad7 level ( < 0.05). The miR-195 inhibitors blocked the activation of HSCs, reduced the expression of miR-195 and -SMA ( < 0.01), and upregulated the expression of Smad7 ( < 0.05).
CONCLUSION
Collectively, we demonstrated that miRNA-195 activated HSCs by targeting Smad7.
Topics: 3' Untranslated Regions; Animals; Diethylnitrosamine; Disease Models, Animal; Gene Expression Regulation; Genes, Reporter; Hepatic Stellate Cells; Liver Cirrhosis; Male; MicroRNAs; Rats; Rats, Sprague-Dawley; Smad7 Protein; Transforming Growth Factor beta1; Up-Regulation
PubMed: 28929107
DOI: 10.1155/2017/1945631 -
Gamma-irradiated β-glucan modulates signaling molecular targets of hepatocellular carcinoma in rats.Tumour Biology : the Journal of the... Aug 2017β-glucans are one of the most abundant forms of polysaccharides known as biological response modifiers which influence host's biological response and stimulate immune...
β-glucans are one of the most abundant forms of polysaccharides known as biological response modifiers which influence host's biological response and stimulate immune system. Accordingly, this study was initiated to evaluate irradiated β-glucan as a modulator for cellular signaling growth factors involved in the pathogenesis of hepatocellular carcinoma in rats. Hepatocellular carcinoma was induced with 20 mg diethylnitrosamine/kg BW. Rats received daily by gastric gavage 65 mg irradiated β-glucan/kg BW. It was found that treatment of rats with diethylnitrosamine induced hepatic injury and caused significant increase in liver injury markers with a concomitant significant increase in both hepatic oxidative and inflammatory indices: alpha-fetoprotein, interferon gamma, and interleukin 6 in comparison with normal and irradiated β-glucan-treated groups. Western immunoblotting showed a significant increase in the signaling growth factors: extracellular signal-regulated kinase 1 and phosphoinositide 3-kinase proteins in a diethylnitrosamine-treated group while both preventive and therapeutic irradiated β-glucan treatments recorded significant improvement versus diethylnitrosamine group via the modulation of growth factors that encounters hepatic toxicity. The transcript levels of vascular endothelial growth factor A and inducible nitric oxide synthase genes were significantly higher in the diethylnitrosamine-treated group in comparison with controls. Preventive and therapeutic treatments with irradiated β-glucan demonstrated that the transcript level of these genes was significantly decreased which demonstrates the protective effect of β-glucan. Histological investigations revealed that diethylnitrosamine treatment affects the hepatic architecture throughout the significant severe appearance of inflammatory cell infiltration in the portal area and congestion in the portal vein in association with severe degeneration and dysplasia in hepatocytes all over hepatic parenchyma. The severity of hepatic architecture changes was significantly decreased with both β-glucan therapeutic and preventive treatments. In conclusion, irradiated β-glucan modulated signal growth factors, vascular endothelial growth factor A, extracellular signal-regulated kinase 1, and phosphatidylinositol-3-kinase, which contributed to experimental hepatocarcinogenesis.
Topics: Animals; Carcinoma, Hepatocellular; Diethylnitrosamine; Disease Models, Animal; Gamma Rays; Humans; Liver; Liver Neoplasms; Mitogen-Activated Protein Kinase 3; Phosphatidylinositol 3-Kinases; Rats; Signal Transduction; Vascular Endothelial Growth Factor A; beta-Glucans
PubMed: 28810822
DOI: 10.1177/1010428317708703 -
Hepatology Communications Nov 2022Phospholipase C gamma 1 (PLCγ1) plays an oncogenic role in several cancers, alongside its usual physiological roles. Despite studies aimed at identifying the effect of...
Phospholipase C gamma 1 (PLCγ1) plays an oncogenic role in several cancers, alongside its usual physiological roles. Despite studies aimed at identifying the effect of PLCγ1 on tumors, the pathogenic role of PLCγ1 in the tumorigenesis and development of hepatocellular carcinoma (HCC) remains unknown. To investigate the function of PLCγ1 in HCC, we generated hepatocyte-specific PLCγ1 conditional knockout (PLCγ1 ; Alb-Cre) mice and induced HCC with diethylnitrosamine (DEN). Here, we identified that hepatocyte-specific PLCγ1 deletion effectively prevented DEN-induced HCC in mice. PLCγ1 ; Alb-Cre mice showed reduced tumor burden and tumor progression, as well as a decreased incidence of HCC and less marked proliferative and inflammatory responses. We also showed that oncogenic phenotypes such as repressed apoptosis, and promoted proliferation, cell cycle progression and migration, were induced by PLCγ1. In terms of molecular mechanism, PLCγ1 regulated the activation of signal transducer and activator of transcription 3 (STAT3) signaling. Moreover, PLCγ1 expression is elevated in human HCC and correlates with a poor prognosis in patients with HCC. Our results suggest that PLCγ1 promotes the pathogenic progression of HCC, and PLCγ1/STAT3 axis was identified as a potential therapeutic target pathway for HCC.
Topics: Humans; Mice; Animals; STAT3 Transcription Factor; Carcinoma, Hepatocellular; Diethylnitrosamine; Liver Neoplasms; Phospholipase C gamma; Cell Proliferation; Carcinogenesis
PubMed: 36153805
DOI: 10.1002/hep4.2077 -
Environmental Science and Pollution... Apr 2023N-nitrosamines are widespread in various bodies of water, which is of great concern due to their carcinogenic risks and harmful mutagenic effects. Livestock rearing,...
N-nitrosamines are widespread in various bodies of water, which is of great concern due to their carcinogenic risks and harmful mutagenic effects. Livestock rearing, domestic, agricultural, and industrial wastewaters are the main sources of N-nitrosamines in environmental water. However, information on the amount of N-nitrosamines these different wastewaters contribute to environmental water is scarce. Here, we investigated eight N-nitrosamines and assessed their mass loadings in the Desheng River to quantify the contributions discharged from different anthropogenic activities. N-nitrosodimethylamine (NDMA) (< 1.6-18 ng/L), N-nitrosomethylethylamine (NMEA) (< 2.2 ng/L), N-nitrosodiethylamine (NDEA) (< 1.7-2.4 ng/L), N-nitrosopyrrolidine (NPYR) (< 1.8-18 ng/L), N-nitrosomorpholine (NMOR) (< 2.0-3.5 ng/L), N-nitrosopiperidine (NPIP) (< 2.2-2.5 ng/L), and N-nitrosodi-n-butylamine (NDBA) (< 3.3-16 ng/L) were detected. NDMA and NDBA were the dominant compounds contributing 89% and 92% to the total N-nitrosamine concentrations. The mean cumulative concentrations of N-nitrosamines in the livestock rearing area (26 ± 11 ng/L) and industrial area (24 ± 4.8 ng/L) were higher than those in the residential area (16 ± 6.3 ng/L) and farmland area (15 ± 5.1 ng/L). The mean concentration of N-nitrosamines in the tributaries (22 ng/L) was slightly higher than that in the mainstem (17 ng/L), probably due to the dilution effect of the mainstem. However, the mass loading assessment based on the river's flow and water concentrations suggested the negligible mass emission of N-nitrosamines into the mainstem from tributaries, which could be due to the small water flow of tributaries. The average mass loads of N-nitrosamines discharged into the mainstem were ranked as the livestock rearing area (742.7 g/d), industrial area (558.6 g/d), farmland area (93.9 g/d), and residential areas (83.2 g/d). In the livestock rearing, residential, and industrial area, NDMA (60.9%, 53.6%, and 46.7%) and NDBA (34.6%, 33.3%, and 44.9%) contributed the most mass loads; NDMA (23.4%), NDEA (15.8%), NPYR (10.1%), NPIP (12.8%), and NDBA (37.8%) contributed almost all the mass loads in the farmland area. Photodegradation amounts of NDMA (0.65 ~ 5.25 µg/(m·day)), NDBA (0.37 ~ 0.91 µg/(m·day)), and NDEA (0 ~ 0.66 µg/(m·day)) were also calculated according to the mass loading. Quantifying the contribution of different anthropogenic activities to the river will provide important information for regional river water quality protection. Risk quotient (RQ) values showed the negligible ecological risks for fish, daphnid, and green algae.
Topics: Wastewater; Rivers; Anthropogenic Effects; Nitrosamines; Diethylnitrosamine; Butylamines
PubMed: 36973615
DOI: 10.1007/s11356-023-26458-8 -
The Journal of Experimental Medicine May 2017Human hepatocellular carcinomas (HCCs), which arise on a background of chronic liver damage and inflammation, express c-Fos, a component of the AP-1 transcription...
Human hepatocellular carcinomas (HCCs), which arise on a background of chronic liver damage and inflammation, express c-Fos, a component of the AP-1 transcription factor. Using mouse models, we show that hepatocyte-specific deletion of c-Fos protects against diethylnitrosamine (DEN)-induced HCCs, whereas liver-specific c-Fos expression leads to reversible premalignant hepatocyte transformation and enhanced DEN-carcinogenesis. c-Fos-expressing livers display necrotic foci, immune cell infiltration, and altered hepatocyte morphology. Furthermore, increased proliferation, dedifferentiation, activation of the DNA damage response, and gene signatures of aggressive HCCs are observed. Mechanistically, c-Fos decreases expression and activity of the nuclear receptor LXRα, leading to increased hepatic cholesterol and accumulation of toxic oxysterols and bile acids. The phenotypic consequences of c-Fos expression are partially ameliorated by the anti-inflammatory drug sulindac and largely prevented by statin treatment. An inverse correlation between c-FOS and the LXRα pathway was also observed in human HCC cell lines and datasets. These findings provide a novel link between chronic inflammation and metabolic pathways important in liver cancer.
Topics: Animals; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Cholesterol; Diethylnitrosamine; Disease Models, Animal; Drosophila Proteins; Liver; Liver Neoplasms; Mice; Proto-Oncogene Proteins c-fos; Repressor Proteins
PubMed: 28356389
DOI: 10.1084/jem.20160935 -
International Journal of Oncology Jan 2017Despite considerable advances in understanding hepatocellular carcinoma, it is one of the common and deadliest cancers worldwide. Hence, increasing efforts are needed...
Despite considerable advances in understanding hepatocellular carcinoma, it is one of the common and deadliest cancers worldwide. Hence, increasing efforts are needed for early diagnosis and effective treatments. Saffron has been recently found to inhibit growth of liver cancer in rats. The aim of this study was to develop an effective method for treatment of liver cancer using magnetite nanoparticles (MNPs) coated with crocin, the main active component of saffron. MNPs were prepared and initially coated with dextran and a cross-linker to enhance conjugation of crocin using a modified coprecipitation method. Cultured HepG2 cells and diethylnitrosamine-injected mice were treated with corcin-coated MNPs and analyzed using cell proliferation assay and immunohistochemical analysis, respectively. Treatment of HepG2 cells with crocin-coated MNPs led to a significant inhibition of their growth as compared to control or those treated with free crocin or uncoated MNPs. Histological examinations of the livers of diethylnitrosamine-injected mice revealed several precancerous changes: multiple proliferative hepatic foci, hyper- or dysplastic transformations of bile ducts/ductules, and nuclear atypia associated with polyploidy, karyomegaly, and vacuolation. Immunohistochemistry using antibodies specific for cell proliferation (Ki-67) and apoptosis (M30-CytoDEATH and Bcl-2) revealed their upregulation during development of precancerous lesions. Using antibodies specific for inflammation (cyclooxygenase-2), oxidative stress (glutathione) and angiogenesis (vascular endothelial growth factor) indicated the involvement of multiple signaling pathways in the development of precancerous lesions. Treatment with crocin-coated MNPs was associated with regression of precancerous lesions, significant upregulation of apoptotic cells and downregulation of Bcl-2 labeling and markers of cell proliferation, inflammation, oxidative stress and angiogenesis. In conclusion, crocin-coated MNPs are more effective than free corcin for treatment of liver precancerous lesions in mice. These findings will help to develop new modalities for early detection and treatment of liver precancerous lesions.
Topics: Animals; Carcinoma, Hepatocellular; Carotenoids; Cell Proliferation; Diethylnitrosamine; Drug Delivery Systems; Hep G2 Cells; Humans; Liver Neoplasms; Magnetite Nanoparticles; Mice; Neoplasms, Experimental; Oxidative Stress
PubMed: 27878253
DOI: 10.3892/ijo.2016.3769 -
Integrative Cancer Therapies Nov 2015Hepatocellular carcinoma (HCC) is one of the common cancers and lethal diseases worldwide. Both oxidative stress and chronic inflammation contribute to the pathogenesis...
Hepatocellular carcinoma (HCC) is one of the common cancers and lethal diseases worldwide. Both oxidative stress and chronic inflammation contribute to the pathogenesis of HCC. Because of limited treatment options and a grave prognosis of HCC, preventive management has been emphasized. The marine macroalgae Ulva lactuca (Ulvaceae) is consumed by humans and livestock because of its nutritional value. Recent studies showed that various extracts of U. lactuca possess antiviral, antiplasmodial, antinephrotoxic, antioxidant, and anti-inflammatory properties. However, very limited information is available on anticancer potential of U. lactuca with no reports on liver cancer chemopreventive efficacy of this marine algae. Accordingly, the present study was initiated to evaluate the possible antihepatocarcinogenic effects and antioxidant mechanisms of action of various U. lactuca extracts against a clinically relevant rodent model of HCC. Initiation of hepatocarcinogenesis was performed in Sprague-Dawley rats by a single injection of dietary carcinogen diethylnitrosamine (DENA, 200 mg/kg, intraperitoneally), followed by promotion with phenobarbital (0.05%) in drinking water. The rats were fed with daily oral dose (50 mg/kg) of polysaccharide sulfate or aqueous extract of U. lactuca for 2, 12, and 24 weeks. At these timepoints, blood samples were taken to measure hepatic injury markers, including alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyl transferase, and bilirubin. The liver tissue was harvested for measurement of hepatic oxidative indices, including lipid peroxidation, reduced glutathione, nitric oxide, catalase, superoxide dismutase, glutathione reductase, and glutathione S-transferase. Hepatic histopathology, immunohistochemical analysis of cell proliferation and apoptosis by DNA fragmentation assay were performed. Our results clearly indicate that sulfated polysaccharides of U. lactuca exert a marked chemoprevention of DENA-initiated hepatocarcinogenesis through inhibition of abnormal cell proliferation and induction of apoptosis. A modest inhibition rat liver carcinogenesis was observed with the aqueous extract. The sulfated polysaccharides altered serum parameters of hepatic damage and modulated various components of the hepatic enzymatic and nonenzymatic antioxidant defense systems. The sulfated polysaccharides from U. lactuca may have unique properties of providing protection against DENA-induced oxidative stress which could contribute to chemoprevention of experimental hepatocarcinogenesis. U. lactuca sulfated polysaccharides could be developed as chemopreventive and therapeutic drug against human HCC.
Topics: Animals; Anticarcinogenic Agents; Apoptosis; Carcinoma, Hepatocellular; Cell Proliferation; Diethylnitrosamine; Liver Neoplasms, Experimental; Male; Oxidative Stress; Phenobarbital; Plant Extracts; Polysaccharides; Rats; Rats, Sprague-Dawley; Time Factors; Ulva
PubMed: 26130745
DOI: 10.1177/1534735415590157 -
Applied Biochemistry and Biotechnology Apr 2023Hepatocellular carcinoma is the second most cause of death among the various cancers worldwide. Recent research searching an alternative therapy for cancer treatment...
Hepatocellular carcinoma is the second most cause of death among the various cancers worldwide. Recent research searching an alternative therapy for cancer treatment without or less side effects. Many studies indicated the beneficial effects of Enhalus acoroides. There has been no scientific validation on antioxidant and chemopreventive potential of ethanolic extract E. acoroides against hepatoma. To assess the hepatoprotective activity of E. acoroides (EEEA) against DEN-induced hepatoma using Wistar albino rats. Animals were distributed into five groups, each containing six rats. To Group I - control rats - normal saline given. Groups II, III, IV and V rats were injection of DEN at a dose of 100 mg/kg body weight i.p. to induce liver cancer. At the commencement of 6th week, Group III rats supplemented with EEEA at a dose of 200 mg/kg body weight/day upto 16 weeks. Group IV rats supplemented with EEEA for 1 week before the administration of DEN and continued till the sixteenth week. Group V supplementation of silymarin at a dose of 100 mg/kg body weight at the beginning of 6th week after the injection of DEN and continued upto 16 weeks and considered as positive control rats. The efficiency of E. acoroides for its antioxidant hepatoprotective and activity evaluated in rats against DEN-induced liver damage. The hepatoprotective ability of EEEA at a dose of 200 mg/kg was examined against DEN at a dose of 100 mg/kg/b.w. induced hepatotoxicity and analysed by evaluating serum liver and kidney marker levels, lipid profile (TG, HDL, LDL and total cholesterol) and serum tumour markers (DNA, RNA, AFP and CEA). Supplementation of EEEA to DEN treated rats was determined by evaluating various antioxidant biomarkers (SOD, CAT, GPx, GSH, Vit E and Vit C). Histopathological studies and morphometric gross analysis were also support the consequences of this study. A significant improvement of antioxidant defence and declined MDA levels within the serum of EEEA treated animals compared to the DEN-induced hepatoma. The supplementation of EEEA declined the serum liver, kidney and serum tumour marker levels and lipid profile as comparatively to Group I rats. The histopathological changes were changed on supplementation of EEEA demonstrating its protecting effects on hepatocytes as comparatively to Group I rats. Our significances recognized that crude extract (ethanol) of E. acoroides revealed a potential impact against DEN-induced hepatoma and assists as a superior choice for chemopreventive treatments.
Topics: Animals; Rats; Carcinoma, Hepatocellular; Antioxidants; Diethylnitrosamine; Liver Neoplasms; Rats, Wistar; Liver; Body Weight; Lipids
PubMed: 35106713
DOI: 10.1007/s12010-022-03832-9 -
Cancer Prevention Research... Jun 2020Hepatocellular carcinoma (HCC) is a highly morbid condition with lack of effective treatment options. HCC arises from chronically inflamed and damaged liver tissue;...
Hepatocellular carcinoma (HCC) is a highly morbid condition with lack of effective treatment options. HCC arises from chronically inflamed and damaged liver tissue; therefore, chemoprevention may be a useful strategy to reduce HCC incidence. Several reports suggest that epigallocatechin gallate (EGCG), extracted from green tea, can suppress liver inflammation and fibrosis in animal models, but its role in HCC chemoprevention is not well established. In this study, male Wistar rats were injected with diethylnitrosamine at 50 mg/kg for 18 weeks to induce cirrhosis and HCC, and EGCG was given in drinking water at a concentration of 0.02%. Clinically achievable dosing of EGCG was well-tolerated in diethylnitrosamine-injured rats and was associated with improved serum liver markers including alanine transaminase, aspartate transaminase, and total bilirubin, and reduced HCC tumor formation. Transcriptomic analysis of diethylnitrosamine-injured hepatic tissue was notable for increased expression of genes associated with the Hoshida high risk HCC gene signature, which was prevented with EGCG treatment. EGCG treatment also inhibited fibrosis progression, which was associated with inactivation of hepatic stellate cells and induction of the senescence-associated secretory phenotype. In conclusion, EGCG administered at clinically safe doses exhibited both chemopreventive and antifibrotic effects in a rat diethylnitrosamine liver injury model.
Topics: Animals; Anticarcinogenic Agents; Biomarkers; Carcinoma, Hepatocellular; Catechin; Cellular Senescence; Diethylnitrosamine; Hepatic Stellate Cells; Liver Cirrhosis; Liver Neoplasms, Experimental; Male; Phenotype; Precancerous Conditions; Rats; Tea; Transcriptome
PubMed: 32253266
DOI: 10.1158/1940-6207.CAPR-19-0383 -
Nanotheranostics 2019: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy, and its current management relies heavily on locoregional therapy for curative therapy,...
: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy, and its current management relies heavily on locoregional therapy for curative therapy, bridge to transplant, and palliative therapy. Locoregional therapies include ablation and hepatic artery therapies such as embolization and radioembolization. In this study we evaluate the feasibility of using novel antivascular ultrasound (AVUS) as a noninvasive locoregional therapy to reduce perfusion in HCC lesions in a rat model and, monitor the effect with contrast-enhanced ultrasound imaging. HCC was induced in 36 Wistar rats by the ingestion of 0.01% diethylnitrosamine (DEN) for 12 weeks. Two therapy regimens of AVUS were evaluated. A primary regimen (n = 19) utilized 2-W/cm, 3-MHz ultrasound (US) for 6 minutes insonation with 0.7 ml of microbubbles administered as an intravenous bolus. An alternate dose at half the primary intensity, sonication time, and contrast concentration was evaluated in 11 rats to assess the efficacy of a reduced dose. A control group (n = 6) received a sham therapy. Tumor perfusion was measured before and after AVUS with nonlinear contrast ultrasound (NLC) and power Doppler (PD). The quantitative perfusion measures included perfusion index (PI), peak enhancement (PE), time to peak (TTP), and perfusion area from NLC and PD scans. Total tumor area perfused during the scan was measured by a postprocessing algorithm called delta projection. Tumor histology was evaluated for signs of tissue injury and for vascular changes using CD31 immunohistochemistry. DEN exposure induced autochthonous hepatocellular carcinoma lesions in all rats. Across all groups prior to therapy, there were no significant differences in the nonlinear contrast observations of peak enhancement and perfusion index. In the control group, there were no significant differences in any of the parameters after sham treatment. After the primary AVUS regimen, there were significant changes in all parameters (p ≤ 0.05) indicating substantial decreases in tumor perfusion. Peak enhancement in nonlinear contrast scans showed a 37.9% ± 10.1% decrease in tumor perfusion. Following reduced-dose AVUS, there were no significant changes in perfusion parameters, although there was a trend for the nonlinear contrast observations of peak enhancement and perfusion index to increase. This study translated low-intensity AVUS therapy into a realistic model of HCC and evaluated its effects on the tumor vasculature. The primary dose of AVUS tested resulted in significant vascular disruption and a corresponding reduction in tumor perfusion. A reduced dose of AVUS, on the other hand, was ineffective at disrupting perfusion but demonstrated the potential for enhancing tumor blood flow. Theranostic ultrasound, where acoustic energy and microbubbles are used to monitor the tumor neovasculature as well as disrupt the vasculature and treat lesions, could serve as a potent tool for delivering noninvasive, locoregional therapy for hepatocellular carcinoma.
Topics: Alkylating Agents; Animals; Blood Vessels; Carcinoma, Hepatocellular; Contrast Media; Diethylnitrosamine; Liver Neoplasms; Male; Microbubbles; Rats; Rats, Wistar; Ultrasonic Therapy; Ultrasonography, Doppler
PubMed: 31687321
DOI: 10.7150/ntno.39514