The Na +/H + exchanger controls deoxycholic acid-induced apoptosis by a H +-activated, Na +-dependent ionic shift in esophageal cells

Aaron Goldman, Hwu Dau Rw Chen, Mohammad R. Khan, Heather Roesly, Kimberly A. Hill, Mohammad - Shahidullah, Amritlal Mandal, Nicholas A Delamere, Katerina Dvorak

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish (US)
Article numbere23835
JournalPLoS One
Volume6
Issue number8
DOIs
StatePublished - 2011

Fingerprint

deoxycholic acid
Sodium-Hydrogen Antiporter
Deoxycholic Acid
apoptosis
Apoptosis
bile acids
Bile Acids and Salts
Amiloride
cells
Acidification
Chemical activation
Cell death
caspase-3
Caspase 3
carcinogenesis
acidification
cell death
Carcinogenesis
Cell Death
Cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

The Na +/H + exchanger controls deoxycholic acid-induced apoptosis by a H +-activated, Na +-dependent ionic shift in esophageal cells. / Goldman, Aaron; Chen, Hwu Dau Rw; Khan, Mohammad R.; Roesly, Heather; Hill, Kimberly A.; Shahidullah, Mohammad -; Mandal, Amritlal; Delamere, Nicholas A; Dvorak, Katerina.

In: PLoS One, Vol. 6, No. 8, e23835, 2011.

Research output: Contribution to journalArticle

Goldman, Aaron ; Chen, Hwu Dau Rw ; Khan, Mohammad R. ; Roesly, Heather ; Hill, Kimberly A. ; Shahidullah, Mohammad - ; Mandal, Amritlal ; Delamere, Nicholas A ; Dvorak, Katerina. / The Na +/H + exchanger controls deoxycholic acid-induced apoptosis by a H +-activated, Na +-dependent ionic shift in esophageal cells. In: PLoS One. 2011 ; Vol. 6, No. 8.
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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.",
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AU - Goldman, Aaron

AU - Chen, Hwu Dau Rw

AU - Khan, Mohammad R.

AU - Roesly, Heather

AU - Hill, Kimberly A.

AU - Shahidullah, Mohammad -

AU - Mandal, Amritlal

AU - Delamere, Nicholas A

AU - Dvorak, Katerina

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AB - 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.

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