Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing β-cells

  • Authors:
    • Jai Parkash
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  • Published online on: October 27, 2010     https://doi.org/10.3892/etm.2010.161
  • Pages: 21-26
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Abstract

We previously showed that tumor necrosis factor-α (TNF-α) induces the dysregulation of intracellular calcium [Ca2+]i in β-cells by decreasing the levels of the cytoplasmic Ca2+ binding protein calbindin-D28k. The purpose of the present study was to test the hypothesis that TNF-α-induced dysregulation of [Ca2+]i in insulin-producing β-cells causes proteolytic degradation of IκBα and consequently leads to the transcriptional activation of nuclear factor-κB (NF-κB). To test this hypothesis, rat insulinoma (RINr 1046-38) cells, which are an insulin-secreting transformed β-cell line that constitutively expresses calbindin-D28k, were treated with increasing concentrations of TNF-α. Using the FunctionELISA procedure to measure degradation of the IκBα subunit as Phospho-IκBα, it was found that, while in the control RIN cell lysate there was no Phospho-IκBα present, in the RIN cells exposed to 2, 5, 10, 20 and 30 ng/ml TNF-α, 17.176±2.85, 17.292±4.35, 53.77±5.63, 30.58±4.89 and 12±3.27 ng/ml Phospho-IκBα/mg of total cell protein was observed, respectively (n=3, P<0.05). Upon treatment of RIN cells with 2, 5, 10, 20 and 30 ng/ml TNF-α, the relative increases in the NF-κB transcriptional activities based on the DNA binding activity of NF-κB determined using an ELISA-based kit were 6.86±0.76-, 8.42±1.27-, 7.8±2.32-, 10.28±1.96- and 6.3±1.57-fold, respectively (n=3, P<0.05). The nuclear translocation of NF-κB measured by immunofluorescence showed that, while the ratio of fluorescence in nuclei to that in the cytoplasm of untreated RIN cells was 0.2078±0.0778 (n=11), in RIN cells treated with 10 ng/ml TNF-α, the ratio was 0.6267±0.1186 (n=11), indicating a statistically significant increase (P<0.05) in the nuclear translocation of NF-κB. These observations suggest that, in insulin-producing β-cells, the TNF-α-induced degradation of IκBα leads to nuclear translocation and the transcriptional activation of NF-κB.
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January-February 2011
Volume 2 Issue 1

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Spandidos Publications style
Parkash J: Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing β-cells. Exp Ther Med 2: 21-26, 2011
APA
Parkash, J. (2011). Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing β-cells. Experimental and Therapeutic Medicine, 2, 21-26. https://doi.org/10.3892/etm.2010.161
MLA
Parkash, J."Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing β-cells". Experimental and Therapeutic Medicine 2.1 (2011): 21-26.
Chicago
Parkash, J."Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing β-cells". Experimental and Therapeutic Medicine 2, no. 1 (2011): 21-26. https://doi.org/10.3892/etm.2010.161