Involvement of endoplasmic reticulum in glycochenodeoxycholic acid-induced apoptosis in rat hepatocytes.

Department of Pharmacology, School of Medicine, Showa University, Hatanodai 1-5-8, Shinagawa-Ku, Tokyo 142-8555, Japan.
Toxicology Letters (Impact Factor: 3.15). 11/2006; 166(2):140-9. DOI: 10.1016/j.toxlet.2006.06.006
Source: PubMed

ABSTRACT In chronic cholestatic liver diseases, accumulation of hydrophobic bile acids is thought to damage hepatocytes. The mechanism of how cells die has been an open debate, but apoptotic pathways are known to involve activation of death receptors and mitochondrial dysfunction. Recently apoptosis via an endoplasmic reticulum (ER) stress-mediated pathway was also found. In this study, we examined whether ER stress is induced in rat hepatocytes by treatment with glycochenodeoxycholic acid (GCDCA, 50-300microM for 1-24h), and if so, whether ER stress-mediated apoptosis occurs in this system. We determined mobility of intracellular calcium ion, activities of calpain and caspase-12, specific to ER stress-mediated apoptosis, and Bip and Chop mRNA expressions, biomarkers of ER stress. We found that GCDCA induces ER-related calcium release within about ten seconds. Significant increases in activities of calpain and caspase-12 were observed after 15h of GCDCA treatment. Bip and Chop mRNA expressions were increased with the treated GCDCA dose and incubation time. Cytochrome c release from mitochondria peaked in about 2h of incubation. These results suggest that ER stress is actually induced by GCDCA, though its role in hepatocellular apoptosis may be smaller than mitochondria-mediated pathway. The presence of ER stress might be important in pathogenesis of cholestatic liver diseases.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Apoptosis resistance is a hallmark of cancer cells. Typically, bile acids induce apoptosis. However during gastrointestinal (GI) tumorigenesis the cancer cells develop resistance to bile acid-induced cell death. To understand how bile acids induce apoptosis resistance we first need to identify the molecular pathways that initiate apoptosis in response to bile acid exposure. In this study we examined the mechanism of deoxycholic acid (DCA)-induced apoptosis, specifically the role of Na(+)/H(+) exchanger (NHE) and Na(+) influx in esophageal cells. In vitro studies revealed that the exposure of esophageal cells (JH-EsoAd1, CP-A) to DCA (0.2 mM-0.5 mM) caused lysosomal membrane perturbation and transient cytoplasmic acidification. Fluorescence microscopy in conjunction with atomic absorption spectrophotometry demonstrated that this effect on lysosomes correlated with influx of Na(+), subsequent loss of intracellular K(+), an increase of Ca(2+) and apoptosis. However, ethylisopropyl-amiloride (EIPA), a selective inhibitor of NHE, prevented Na(+), K(+) and Ca(2+) changes and caspase 3/7 activation induced by DCA. Ouabain and amphotericin B, two drugs that increase intracellular Na(+) levels, induced similar changes as DCA (ion imbalance, caspase3/7 activation). On the contrary, DCA-induced cell death was inhibited by medium with low a Na(+) concentrations. In the same experiments, we exposed rat ileum ex-vivo to DCA with or without EIPA. Severe tissue damage and caspase-3 activation was observed after DCA treatment, but EIPA almost fully prevented this response. In summary, NHE-mediated Na(+) influx is a critical step leading to DCA-induced apoptosis. Cells tolerate acidification but evade DCA-induced apoptosis if NHE is inhibited. Our data suggests that suppression of NHE by endogenous or exogenous inhibitors may lead to apoptosis resistance during GI tumorigenesis.
    PLoS ONE 01/2011; 6(8):e23835. · 3.53 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The endoplasmic reticulum (ER) as an intracellular Ca(2+) store not only sets up cytosolic Ca(2+) signals, but, among other functions, also assembles and folds newly synthesized proteins. Alterations in ER homeostasis, including severe Ca(2+) depletion, are an upstream event in the pathophysiology of many diseases. On the one hand, insufficient release of activator Ca(2+) may no longer sustain essential cell functions. On the other hand, loss of luminal Ca(2+) causes ER stress and activates an unfolded protein response, which, depending on the duration and severity of the stress, can reestablish normal ER function or lead to cell death. We will review these various diseases by mainly focusing on the mechanisms that cause ER Ca(2+) depletion.
    Cold Spring Harbor perspectives in biology 03/2011; 3(6). · 9.63 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Oesophageal adenocarcinoma (OA) incidence is rising and prognosis is poor. Understanding the molecular basis of this malignancy is key to finding new prevention and treatment strategies. Gastroesophageal reflux disease (GORD) is the primary cause of OA, usually managed with acid suppression therapy. However, this often does little to control carcinogenic bile acid reflux. The transcription factor nuclear factor kappa B (NF-κB) plays a key role in the pathogenesis of OA and its activity is associated with a poor response to chemotherapy, making it an attractive therapeutic target. We sought to decipher the role of different bile acids in NF-κB activation in oesophageal cell lines using short, physiologically relevant exposure times. The effect of an acidic or neutral extracellular pH was investigated concurrently, to mimic in vivo conditions associated with or without acid suppression. We found that some bile acids activated NF-κB to a greater extent when combined with acid, whereas others did so in its absence, at neutral pH. The precise composition of an individual's reflux, coupled with whether they are taking acid suppressants may therefore dictate the extent of NF-κB activation in the oesophagus, and hence the likelihood of histological progression and chemotherapy success. Regardless of pH, the kinase IKK was pivotal in mediating reflux induced NF-κB activation. Its importance was confirmed further as its increased activation was associated with histological progression in patient samples. We identified further kinases important in acid or bile induced NF-κB signaling in oesophageal cells, which may provide suitable targets for therapeutic intervention. © 2014 Wiley Periodicals, Inc.
    International Journal of Cancer 06/2014; · 6.20 Impact Factor