Characterizing the differential physiological effects of adipocytokines visfatin and resistin in liver cancer cells

2019 ◽  
Vol 38 (2) ◽  
Author(s):  
Candace Miethe ◽  
Megan Zamora ◽  
Linda Torres ◽  
Kelsie G. Raign ◽  
Curissa J. Groll ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Liver Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Fatty Acid Synthase ◽  
Liver Tumors ◽  
Health Concern ◽  
Liver Cancer Cells ◽  
Erk Phosphorylation ◽  
Huh7 Cells

Abstract Background Obesity, a major public health concern, increases the risk of developing liver cancer which is the leading cause of cancer-related deaths worldwide. Obesity is associated with increased adiposity and macrophage infiltration both of which promote secretion of adipokines and cytokines in the tumor microenvironment. Specifically, visfatin and resistin have been detected at higher levels in the serum of obese individuals and liver tumors. However, the contribution of these adipocytokines in the progression of liver cancer remains unclear. Materials and methods The objective of this study was to characterize the effects of visfatin and resistin on HepG2, SNU-449 and HuH7 liver cancer cells. Cells exposed to visfatin and resistin were analyzed for fatty acid synthase protein, and phosphorylation of Akt and ERK tumorigenic signaling pathways, cell viability, lipogenesis, reactive oxygen species (ROS), matrix metallopeptidase 9 (MMP-9) enzyme activity and invasion. Results HepG2, SNU-449, and HuH7 liver cancer cells treated with visfatin and resistin increased cell viability, invasion, FASN protein, and Akt and ERK phosphorylation. Visfatin and resistin selectively increased ROS production in HepG2 and SNU-449 cells while there was no statistical difference in HuH7 cells. Visfatin and resistin stimulated lipogenesis in HepG2 cells while visfatin increased lipogenesis in SNU-449 cells, and visfatin nor resistin had an effect on lipogenesis in HuH7 cells. Lastly, visfatin and resistin increased MMP-9 enzyme activity in HepG2 and HuH-7 cells but only visfatin increased MMP-9 activity in SNU-449 cells. Conclusions Future studies are needed to determine if inhibition of ERK and Akt suppresses the visfatin and resistin-induced invasive liver cancer phenotype.

Inhibition of PI3K/Akt and ERK signaling decreases visfatin-induced invasion in liver cancer cells

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Candace Miethe ◽  
Linda Torres ◽  
Megan Zamora ◽  
Ramona S. Price
Keyword(s):  
Liver Cancer ◽  
Cancer Cells ◽  
Signaling Pathways ◽  
Fatty Acid Synthase ◽  
Erk Signaling ◽  
Ros Production ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Liver Cancer Cells ◽  
Phosphorylated Akt ◽  
Erk Inhibition

Abstract Objectives Visfatin is found in adipose tissue and is referred to as nicotinamide phosphoribosyltransferase (Nampt). Visfatin has anti-apoptotic, proliferative, and metastatic properties and may mediate its effects via ERK and PI3K/Akt signaling. Studies have yet to determine whether inhibition of kinase signaling will suppress visfatin-induced liver cancer. The purpose of this study was to determine which signaling pathways visfatin may promote liver cancer progression. Methods HepG2 and SNU-449 liver cancer cells were exposed to visfatin with or without ERK or PI3K/Akt inhibitor, or both inhibitors combined. These processes that were assessed: proliferation, reactive oxygen species (ROS), lipogenesis, invasion, and matrix metalloproteinase (MMP). Results Inhibition of PI3K/Akt and combination of inhibitors suppressed visfatin-induced viability. ERK inhibition in HepG2 cells decreased visfatin-induced proliferation. ERK inhibitor alone or in combination with PI3K inhibitors effectively suppressed MMP-9 secretion and invasion in liver cancer cells. PI3K and ERK inhibition and PI3K inhibition alone blocked visfatin’s ROS production in SNU-449 cells. These results corresponded with a decrease in phosphorylated Akt and ERK, β-catenin, and fatty acid synthase. Conclusions Akt and ERK inhibition differentially regulated physiological changes in liver cancer cells. Inhibition of Akt and ERK signaling pathways suppressed visfatin-induced invasion, viability, MMP-9 activation, and ROS production.

scholarly journals Ginsenoside Compound K Regulates HIF-1α-Mediated Glycolysis Through Bclaf1 to Inhibit the Proliferation of Human Liver Cancer Cells

2020 ◽  
Vol 11 ◽  
Author(s):  
Silin Zhang ◽  
Meilan Zhang ◽  
Jiaxin Chen ◽  
Jiaqi Zhao ◽  
Jielin Su ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Cancer Cells ◽  
Hepatoma Cells ◽  
Dependent Manner ◽  
Compound K ◽  
Liver Cancer Cells ◽  
Hypoxic Conditions ◽  
Huh7 Cells ◽  
Glycolysis Pathway ◽  
Ginsenoside Compound K

This study aimed to demonstrate that ginsenoside compound K (20 (S)-ginsenoside CK; CK) downregulates Bcl-2-associated transcription factor 1 (Bclaf1), which inhibits the hypoxia-inducible factor-1α (HIF-1α)-mediated glycolysis pathway to inhibit the proliferation of liver cancer cells. Treatment of hepatoma cells (Bel-7404 and Huh7) under hypoxic conditions with different concentrations of CK showed that CK inhibited the proliferation of hepatoma cells in a time- and concentration-dependent manner; furthermore, the ability of the cells to form colonies was reduced, and cell growth was blocked in the G0/G1 phase. CK promoted the degradation of HIF-1α ubiquitination in liver cancer cells by regulating the expression of HIF-1α and related ubiquitination proteins; moreover, it reduced the activity of key enzymes involved in glycolysis, the pressure of cellular glycolysis, and the rate of real-time ATP production, thereby inhibiting the glycolysis pathway. It also decreased the expression of Bclaf1 in hypoxic liver cancer cells and thus reduced the ability of Bclaf1 to bind to HIF-1α. CK treatment of Bel-7404 and Huh7 cells with CRISPR/Cas9-engineered knock out of Bclaf1 gene under hypoxic conditions further suppressed the expression of HIF-1α, promoted HIF-1α ubiquitination, and inhibited the glycolysis pathway. In a rat model of primary liver cancer induced by diethylnitrosamine, positron emission tomography and computed tomography scans showed that after CK administration, tumor tissue volumes were reduced and glucose uptake capacity decreased. Increased Bclaf1 and HIF-1α expression promoted the ubiquitination of HIF-1α and inhibited the glycolysis pathway, thereby inhibiting the proliferation of liver cancer cells. In summary, this study confirmed by in vitro and in vivo experiments that in hypoxic liver cancer cells CK downregulates the expression of Bclaf1, inhibits the HIF-1α-mediated glycolysis pathway, and inhibits cell proliferation, suggesting that the CK-mediated effects on Bclaf1 may represent a novel therapeutic approach for the treatment of liver cancer patients.

scholarly journals Paeoniflorin inhibits cell viability and invasion of liver cancer cells via inhibition of Skp2

2020 ◽  
Author(s):  
Hong Liu ◽  
Lili Zang ◽  
Jun Zhao ◽  
Zhaolin Wang ◽  
Lingyun Li
Keyword(s):  
Liver Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Liver Cancer Cells

Estrogen signalling through amphiregulin may be implicated in human hepatocellular carcinoma

2011 ◽  
Vol 5 (3) ◽  
pp. 153-160 ◽  
Author(s):  
Giuseppe Carruba ◽  
Vitale Miceli ◽  
Letizia Cocciadiferro ◽  
Maurizio Zarcone ◽  
Biagio Agostara ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Liver Cancer ◽  
Cancer Cells ◽  
Human Hepatocellular Carcinoma ◽  
Mrna Levels ◽  
Wild Type ◽  
Liver Cancer Cells ◽  
Huh7 Cells ◽  
Liver Tissues

AbstractBackground:We investigated aromatase (Aro)-driven estrogen formation in non-tumoral and malignant liver tissues and cells, also in relation to expression of the estrogen receptors α and β (ERα and ERβ) and amphiregulin (AREG), aiming to gain insights into the potential role of estrogens in human hepatocellular carcinoma (HCC).Materials and methods:Chromatographic and reverse transcriptase polymerase chain reaction (RT-PCR) analyses were used to assess activity and expression of the Aro enzyme and AREG as well as the expression of wild-type and variant ERs, both in vivo and in vitro.Results:Following 24 h and 72 h incubation of liver tissues or cells with testosterone, human HCC tissues and HepG2 hepatoma cells showed elevated Aro activity (estrogen formation, respectively, of 20% and 52%–99%). By contrast, no Aro activity could be detected in non-tumoral tissues and HA22T liver cancer cells. Cirrhotic samples and Huh7 cells exhibited intermediate enzyme activity, with estrogen formation of 4% and 34%, respectively. Markedly lower or undetectable Aro mRNA levels were observed in HA22T cells and non-tumoral liver tissues compared with HepG2 cells and HCC samples. Cirrhotic specimens displayed variable transcript levels. Interestingly, no or low expression of wild-type ERα and ERβ could be observed in liver cancer cells and malignant tissues. However, ubiquitous expression of the hERα46 variant and occasional expression of the hERβ2/Cx variant were observed in cancer tissues and cells.Conclusions:It is noteworthy that the pattern of wild-type ERα was inversely related to Aro, whilst AREG expression was consistently associated with that of Aro. This combined evidence suggests that locally elevated Aro activity may increase malignant cell proliferation also through AREG signalling.

scholarly journals RRM2 protects against ferroptosis and is a tumor biomarker for liver cancer

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Yueyue Yang ◽  
Jiafei Lin ◽  
Susu Guo ◽  
Xiangfei Xue ◽  
Yikun Wang ◽  
...  
Keyword(s):  
Cell Death ◽  
Liver Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Diagnostic Value ◽  
Regulatory Subunit ◽  
Enzyme Linked Immunosorbent Assay ◽  
Glutathione Synthetase ◽  
Tumor Biomarker ◽  
Liver Cancer Cells

Abstract Background Ferroptosis is the process of cell death triggered by lipid peroxides, and inhibition of glutathione (GSH) synthesis leads to ferroptosis. Liver cancer progression is closely linked to ferroptosis suppression. However, the mechanism by which inhibition of GSH synthesis suppresses potential ferroptosis of liver cancer cells and whether ferroptosis-related liver cancer biomarkers have a promising diagnostic value remain unknown. Methods Ribonucleotide reductase regulatory subunit M2 (RRM2) levels were measured using an enzyme linked immunosorbent assay (ELISA), quantitative RT-PCR (qPCR), immunoblotting (IB) and immunochemistry (IHC). Cell viability and cell death were measured by a CellTiter-Glo luminescent cell viability assay and staining with SYTOX Green followed by flow cytometry, respectively. Metabolites were measured using the indicated kits. The Interaction between glutathione synthetase (GSS) and RRM2 was measured using immunofluorescence (IF), co-immunoprecipitation (co-IP) and the proximal ligation assay (PLA). The diagnostic value was analyzed using the area under the receiver operating characteristic curve (AUC-ROC). Bioinformatics analysis was performed using the indicated database. Results RRM2 showed specifically elevated levels in liver cancer and inhibited ferroptosis by stimulating GSH synthesis via GSS. Mechanistically, phosphorylation of RRM2 at the Threonine 33 residue (T33) was maintained at normal levels to block the RRM2–GSS interaction and therefore protected RRM2 and GSS from further proteasome degradation. However, under ferroptotic stress, RRM2 was dephosphorylated at T33, thus the RRM2–GSS interaction was promoted. This resulted in the translocation of RRM2 and GSS to the proteasome for simultaneous degradation. Clinically, serum RRM2 was significantly associated with serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (γ-GT), albumin (ALB) and total bilirubin. The AUC-ROC for the combination of RRM2 with AFP was 0.947, with a sensitivity of 88.7% and a specificity of 97.0%, which indicates better diagnostic performance compared to either RRM2 or AFP alone. Conclusion RRM2 exerts an anti-ferroptotic role in liver cancer cells by sustaining GSH synthesis. Serum RRM2 will be useful as a biomarker to evaluate the degree to which ferroptosis is suppressed and improve diagnostic efficiency for liver cancer.

scholarly journals A novel small molecule STAT3 inhibitor, LY5, inhibits cell viability, colony formation, and migration of colon and liver cancer cells

Oncotarget ◽  
2016 ◽  
Vol 7 (11) ◽  
pp. 12917-12926 ◽  
Author(s):  
Chongqiang Zhao ◽  
Wenlong Wang ◽  
Wenying Yu ◽  
David Jou ◽  
Yina Wang ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Small Molecule ◽  
Colony Formation ◽  
Liver Cancer Cells ◽  
And Migration

scholarly journals Cell viability in normal fibroblasts and liver cancer cells after treatment with iron (III), nickel (II), and their mixture

2018 ◽  
Vol 62 (4) ◽  
pp. 535-542 ◽  
Author(s):  
Sylwia Terpiłowska ◽  
Dorota Siwicka-Gieroba ◽  
Andrzej Krzysztof Siwicki
Keyword(s):  
Liver Cancer ◽  
Synergistic Effect ◽  
Cell Viability ◽  
Cancer Cells ◽  
Hepg2 Cells ◽  
Cell Metabolism ◽  
Nickel Chloride ◽  
Iron Chloride ◽  
Liver Cancer Cells ◽  
Diet Supplements

Abstract Introduction: Nickel and iron are very commonly occurring metals. Nickel is used in industry, but nowadays it is also used in medical biomaterials. Iron is an element necessary for cell metabolism and is used in diet supplements and biomaterials, whence it may be released along with nickel. Material and Methods: BALB/3T3 and HepG2 cells were incubated with iron chloride or nickel chloride at concentrations ranging from 100 to 1,400 µM. The following mixtures were used: iron chloride 200 µM plus nickel chloride 1,000 µM, or iron chloride 1,000 µM plus nickel chloride 200 µM. The cell viability was determined with MTT, LHD, and NRU tests. Results: A decrease in cell viability was observed after incubating the BALB/3T3 and HepG2 cells with iron chloride or nickel chloride. A synergistic effect was observed after iron chloride 1,000 μM plus nickel chloride 200 μM treatment in all assays. Moreover, the same effect was observed in the pair iron chloride 200 μM plus nickel chloride 1,000 μM in the LDH and NRU assays. Conclusions: Iron (III) and nickel (II) decrease cell viability. Iron chloride at a concentration of 200 µM protects mitochondria from nickel chloride toxicity.

scholarly journals Influence of inosine pranobex on cell viability in normal fibroblasts and liver cancer cells

2018 ◽  
Vol 62 (2) ◽  
pp. 215-220 ◽  
Author(s):  
Sylwia Tobólska ◽  
Sylwia Terpiłowska ◽  
Jerzy Jaroszewski ◽  
Andrzej Krzysztof Siwicki
Keyword(s):  
Liver Cancer ◽  
Cell Membrane ◽  
Cell Viability ◽  
Cancer Cells ◽  
Hepg2 Cells ◽  
Viral Infections ◽  
Inosine Pranobex ◽  
Liver Cancer Cells ◽  
Mtt Reduction

AbstractIntroductionInosine pranobex (Isoprinosine) stimulates cell-mediated immune responses to viral infections in humans and might have also therapeutic use in animals. The aim of this study was to compare three in vitro cytotoxicity assays on mouse embryo fibroblasts and liver cancer cells and determine their ability to detect early cytotoxic effects for inosine pranobex.Material and MethodsBALB/3T3 clone A31and HepG2 cells were incubated with inosine pranobex at concentrations from 0.1 to 1,000 μg/mL. Cell viability was determined with the MTT reduction, the LHD release, and the NRU tests.ResultsA decrease in the cell viability was observed after incubating the BALB/3T3 clone A31and HepG2 cells with inosine pranobex.ConclusionsBased on the cytotoxicity endpoints measured in these investigations in BALB/3T3 clone A31cells, it can be concluded that the cell membrane may be the first part of the cell to be affected by inosine pranobex. The disintegration of lysosomes and mitochondria follows mitochondria damage. In HepG2 cells likewise, the cell membrane may be the first part of the cell to be affected by inosine pranobex. Also in liver cancer cells, the disintegration of mitochondria (assessed with the MTT reduction assay) and next of lysosomes (assessed with the NRU assay) follows mitochondria damage.

Panobinostat treatment determines oncogenic miRNAs down-regulation in liver cancer cells

2012 ◽  
Vol 50 (01) ◽  
Author(s):  
A Henrici ◽  
R Montalbano ◽  
K Quint ◽  
M Ocker ◽  
P Di Fazio
Keyword(s):  
Liver Cancer ◽  
Cancer Cells ◽  
Down Regulation ◽  
Liver Cancer Cells ◽  
Oncogenic Mirnas
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