Polyamines are required for activation of c-Jun NH2-terminal kinase and apoptosis in response to TNF-α in IEC-6 cells

Sujoy Bhattacharya, Ramesh M. Ray, Mary Jane Viar, Leonard R. Johnson

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Abstract

Intracellular polyamine homeostasis is important for the regulation of cell proliferation and apoptosis and is necessary for the balanced growth of cells and tissues. Polyamines have been shown to play a role in the regulation of apoptosis in many cell types, including IEC-6 cells, but the mechanism is not clear. In this study, we analyzed the mechanism by which polyamines regulate the process of apoptosis in response to tumor necrosis factor-α (TNF-α). TNF-α or cycloheximide (CHX) alone did not induce apoptosis in IEC-6 cells. Significant apoptosis was observed when CHX was given along with TNF-α, as indicated by a significant increase in the detachment of cells, caspase-3 activity, and DNA fragmentation. Polyamine depletion by treatment with α-difluoromethylornithine significantly reduced the level of apoptosis, as judged by DNA fragmentation and the caspase-3 activity of attached cells. Apoptosis in IEC-6 cells was accompanied by the activation of upstream caspases-6, -8, and -9 and NH 2-terminal c-Jun kinase (JNK). Inhibition of JNK activation prevented caspase-9 activation. Polyamine depletion prevented the activation of JNK and of caspases-6, -8, -9, and -3. SP-600125, a specific inhibitor of JNK activation, prevented cytochrome c release from mitochondria, JNK activation, DNA fragmentation, and caspase-9 activation in response to TNF-α/CHX. In conclusion, we have shown that polyamine depletion delays and decreases TNF-α-induced apoptosis in IEC-6 cells and that apoptosis is accompanied by the release of cytochrome c, the activation of JNK, and of upstream caspases as well as caspase-3. Polyamine depletion prevented JNK activation, which may confer protection against apoptosis by modulation of upstream caspase-9 activation.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume285
Issue number5 48-5
StatePublished - Nov 2003

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JNK Mitogen-Activated Protein Kinases
Polyamines
Tumor Necrosis Factor-alpha
Apoptosis
Phosphotransferases
Caspase 9
DNA Fragmentation
Cycloheximide
Caspase 6
Caspase 3
Caspase 8
Cytochromes c
Mitogen-Activated Protein Kinase 9
Eflornithine
Caspases
Mitochondria
Homeostasis
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Gastroenterology
  • Physiology

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Polyamines are required for activation of c-Jun NH2-terminal kinase and apoptosis in response to TNF-α in IEC-6 cells. / Bhattacharya, Sujoy; Ray, Ramesh M.; Viar, Mary Jane; Johnson, Leonard R.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 285, No. 5 48-5, 11.2003.

Research output: Contribution to journalArticle

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