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Related Experiment Videos

Proglucagon expression, posttranslational processing and secretion in SV40-transformed islet cells.

K I Shennan1, J J Holst, K Docherty

  • 1Department of Medicine, University of Birmingham, Queen Elizabeth Hospital, U.K.

Molecular and Cellular Endocrinology
|November 1, 1989
PubMed
Summary

A novel hamster islet cell line variant, HIT T15-G, spontaneously evolved to express glucagon. This cell line exhibits regulated glucagon gene expression and secretion, offering insights into pancreatic alpha cell function.

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Area of Science:

  • Endocrinology
  • Cell Biology
  • Molecular Biology

Background:

  • HIT T15 is a hamster islet B cell line.
  • A spontaneously evolved variant, HIT T15-G, now expresses glucagon.

Purpose of the Study:

  • To investigate glucagon gene expression, proglucagon processing, and secretion in HIT T15-G cells.
  • To characterize the regulatory mechanisms of glucagon in this novel cell line.

Main Methods:

  • Forskolin, phorbol ester (TPA), and ionomycin treatments to assess gene expression.
  • Analysis of proglucagon processing products, including GLP-I, GLP-II, and MPF.
  • Stimulation of glucagon secretion using forskolin, adrenalin, arginine, KCl, and glucose.

Main Results:

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  • Glucagon mRNA increased with forskolin but not TPA or ionomycin.
  • Proglucagon processed to glucagon and GLP peptides.
  • Glucagon secretion stimulated by forskolin, adrenalin, arginine, and KCl, but not glucose.

Conclusions:

  • HIT T15-G cells display regulated glucagon expression and secretion.
  • These cells represent a less differentiated form of HIT T15, with a dominant, yet incomplete, alpha cell phenotype.