Expression of the electrogenic Na+-HCO3--cotransporter NBCe1 in tumoral insulin-producing BRIN-BD11 cells

Cell Physiol Biochem. 2009;24(3-4):187-92. doi: 10.1159/000233257. Epub 2009 Aug 3.

Abstract

Background/aims: The expression of the electrogenic Na+-HCO3--cotransporter NBCe1 was recently documented in rat pancreatic islet B-cells, it being speculated that such a protein participates in the extrusion of bicarbonate generated by the oxidative catabolism of nutrients from insulin-producing cells. Considering the prevalence of a Crabtree effect in tumoral insulin-producing cells, the possible presence of NBCe1 was now investigated in BRIN-BD11 cells, an insulin-producing cell line established by electrofusion of normal pancreatic B-cells with immortalized RINm5F cells.

Methods: The possible presence of NBCe1 in BRIN-BD11 cells was investigated by RT-PCR, western blot analysis and immunocytochemistry. The release of insulin and net uptake of 22Na+ were also measured in the BRIN-BD11 cells.

Results: RT-PCR, western blot analysis and immunocytochemistry documented the presence of NBCe1 in BRIN-BD11 cells. A reported inhibitor of NBCe1, i.e. tenidap, (50-100 microM), inhibited basal and hypotonicity-induced insulin release from the BRIN-BD11 cells, whilst increasing the net uptake of 22Na+ by the same cells. The latter effect was, in relative terms, more pronounced in the presence than absence of ouabain.

Conclusion: BRIN-BD11 cells, like normal pancreatic islet B-cells, express NBCe1, with predominance of the B variant of this electrogenic Na+-HCO3--cotransporter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Transformed
  • Cell Line, Tumor
  • Dose-Response Relationship, Drug
  • Fluoresceins / metabolism
  • Fluorescent Antibody Technique, Indirect
  • Fluorescent Dyes / metabolism
  • Gene Expression
  • Glucose / metabolism
  • Immunohistochemistry
  • Indoles / metabolism
  • Indoles / pharmacology
  • Insulin / metabolism*
  • Insulin Secretion
  • Islets of Langerhans / metabolism*
  • Ouabain / pharmacology
  • Oxindoles
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Rats
  • Rats, Inbred Strains
  • Sodium / metabolism
  • Sodium-Bicarbonate Symporters / genetics*
  • Sodium-Bicarbonate Symporters / metabolism
  • Time Factors

Substances

  • Fluoresceins
  • Fluorescent Dyes
  • Indoles
  • Insulin
  • Oxindoles
  • Protein Isoforms
  • Slc4a4 protein, rat
  • Sodium-Bicarbonate Symporters
  • DAPI
  • Ouabain
  • tenidap
  • Sodium
  • Glucose