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The transcription factor PAX4 acts as a survival gene in INS-1E insulinoma cells

Oncogene volume 26, pages 4261–4271 (2007)Cite this article

Abstract

The paired/homeodomain transcription factor Pax4 is essential for islet β-cell generation during pancreas development and their survival in adulthood. High Pax4 expression was reported in human insulinomas indicating that deregulation of the gene may be associated with tumorigenesis. We report that rat insulinoma INS-1E cells express 25-fold higher Pax4 mRNA levels than rat islets. In contrast to primary β-cells, activin A but not betacellulin or glucose induced Pax4 mRNA levels indicating dissociation of Pax4 expression from insulinoma cell proliferation. Short hairpin RNA adenoviral constructs targeted to the paired domain or homeodomain (viPax4PD and viPax4HD) were generated. Pax4 mRNA levels were lowered by 73 and 50% in cells expressing either viPax4PD or viPax4HD. Transcript levels of the Pax4 target gene bcl-xl were reduced by 53 and 47%, whereas Pax6 and Pdx1 mRNA levels were unchanged. viPax4PD-infected cells displayed a twofold increase in spontaneous apoptosis and were more susceptible to cytokine-induced cell death. In contrast, proliferation was unaltered. RNA interference-mediated repression of insulin had no adverse effects on either Pax4 or Pdx1 expression as well as on cell replication or apoptosis. These results indicate that Pax4 is redundant for proliferation of insulinoma cells, whereas it is essential for survival through upregulation of the antiapoptotic gene bcl-xl.

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Abbreviations

BrdU:

5-bromo-2′-deoxy-uridine

BSA:

bovine serum albumin

DAPI, 4′:

6-diamidino-2-phenylindole

FCS:

fetal calf serum

shRNA:

short hairpin RNA

PBS:

phosphate-buffered saline

TUNEL:

terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling

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Acknowledgements

We are grateful to Elodie Husi, Mathurin Baquié and Delphine Chesnel for their expert technical assistance. This work was supported by grants from the Swiss National Science Foundation (#3100A0-107682/1 to BRG) and from the Juvenile Diabetes Research Foundation (#7-2005-1158 to CBW).

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Authors and Affiliations

  1. Department of Cell Physiology and Metabolism, University Medical Center, Geneva 4, Switzerland

    T Brun, D L Duhamel, K H Hu He, C B Wollheim & B R Gauthier

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Correspondence to T Brun.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Brun, T., Duhamel, D., Hu He, K. et al. The transcription factor PAX4 acts as a survival gene in INS-1E insulinoma cells. Oncogene 26, 4261–4271 (2007). https://doi.org/10.1038/sj.onc.1210205

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