Characterization of a cell culture model for clinically aggressive hepatocellular carcinoma induced by chronic hypoxia

Malenstein, Hannah van; Verslype, Chris; Windmolders, Petra; Eijsden, Rudy van; Nevens, Frederik; Pelt, Jos van

doi:10.1016/j.canlet.2011.09.039

Cited by 13 publications

(2 citation statements)

References 40 publications

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“…MIN6 cells used for experiments ranged from passages 22 to 36. INS-1E cells (a kind gift from Prof. C. Wollheim, Centre Medical Universitaire, Geneva, Switzerland, [20]), HepG2 (ATCC HB-8065, Rockville, MD) and COS-1 (ATCC CRL1650, Manassas, VA) cell lines were cultured as described before [21], [22].…”

Section: Methodsmentioning

confidence: 99%

Foodborne Cereulide Causes Beta-Cell Dysfunction and Apoptosis

et al. 2014

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Aims/HypothesisTo study the effects of cereulide, a food toxin often found at low concentrations in take-away meals, on beta-cell survival and function.MethodsCell death was quantified by Hoechst/Propidium Iodide in mouse (MIN6) and rat (INS-1E) beta-cell lines, whole mouse islets and control cell lines (HepG2 and COS-1). Beta-cell function was studied by glucose-stimulated insulin secretion (GSIS). Mechanisms of toxicity were evaluated in MIN6 cells by mRNA profiling, electron microscopy and mitochondrial function tests.Results24 h exposure to 5 ng/ml cereulide rendered almost all MIN6, INS-1E and pancreatic islets apoptotic, whereas cell death did not increase in the control cell lines. In MIN6 cells and murine islets, GSIS capacity was lost following 24 h exposure to 0.5 ng/ml cereulide (P<0.05). Cereulide exposure induced markers of mitochondrial stress including Puma (p53 up-regulated modulator of apoptosis, P<0.05) and general pro-apoptotic signals as Chop (CCAAT/-enhancer-binding protein homologous protein). Mitochondria appeared swollen upon transmission electron microscopy, basal respiration rate was reduced by 52% (P<0.05) and reactive oxygen species increased by more than twofold (P<0.05) following 24 h exposure to 0.25 and 0.50 ng/ml cereulide, respectively.Conclusions/InterpretationCereulide causes apoptotic beta-cell death at low concentrations and impairs beta-cell function at even lower concentrations, with mitochondrial dysfunction underlying these defects. Thus, exposure to cereulide even at concentrations too low to cause systemic effects appears deleterious to the beta-cell.

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Section: Methodsmentioning

confidence: 99%

Foodborne Cereulide Causes Beta-Cell Dysfunction and Apoptosis

et al. 2014

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“…The therapeutic options for advanced or metastatic HCC are very limited [8], partially because the molecular mechanisms underlying hepatocarcinogenesis remain unclear [9]. Suitable animal models and efficient cell culture systems have provided a major breakthrough for understanding these mechanisms [10], [11].…”

Section: Introductionmentioning

confidence: 99%

Overexpression of Endothelin 1 Triggers Hepatocarcinogenesis in Zebrafish and Promotes Cell Proliferation and Migration through the AKT Pathway

Liao

Yang

et al. 2014

PLoS ONE

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Hepatocarcinogenesis commonly involves the gradual progression from hepatitis to fibrosis and cirrhosis, and ultimately to hepatocellular carcinoma (HCC). Endothelin 1 (Edn1) has been identified as a gene that is significantly up-regulated in HBx-induced HCC in mice. In this study, we further investigated the role of edn1 in hepatocarcinogenesis using a transgenic zebrafish model and a cell culture system. Liver-specific edn1 expression caused steatosis, fibrosis, glycogen accumulation, bile duct dilation, hyperplasia, and HCC in zebrafish. Overexpression of EDN1 in 293T cells enhanced cell proliferation and cell migration in in vitro and xenotransplantation assays and was accompanied with up-regulation of several cell cycle/proliferation- and migration-specific genes. Furthermore, expression of the unfolded protein response (UPR) pathway-related mediators, such as spliced XBP1, ATF6, IRE1, and PERK, was also up-regulated at both the RNA and protein levels. In the presence of an EDN1 inhibitor or an AKT inhibitor, these increases were diminished and the EDN1-induced migration ability also was disappeared, suggesting that the EDN1 effects act through activation of the AKT pathway to enhance the UPR and subsequently activate the expression of downstream genes. Additionally, p-AKT is enhanced in the edn1 transgenic fish compared to the GFP-mCherry control. The micro RNA miR-1 was found to inhibit the expression of EDN1. We also observed an inverse correlation between EDN1 and miR-1 expression in HCC patients. In conclusion, our data suggest that EDN1 plays an important role in HCC progression by activating the PI3K/AKT pathway and is regulated by miR-1.

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