TY - JOUR

T1 - Peptide production and secretion in GLUTag, NCI-H716 and STC-1 cells: a comparison to native L-cells.

AU - Kuhre, Rune Ehrenreich

AU - Albrechtsen, Nicolai Jacob Wewer

AU - Deacon, Carolyn F.

AU - Balk-Møller, Emilie

AU - Rehfeld, Jens Frederik

AU - Relman, Frank

AU - Gribble, Fiona Mary

AU - Holst, Jens Juul

PY - 2016

Y1 - 2016

N2 - GLUTag, NCI-H716 and STC-1 are cell lines that are widely used to study mechanisms underlying secretion of glucagon-like peptide (GLP-1), but the extent to which they resemble native L-cells is unknown. We used validated immunoassays for 14 different hormones to analyze peptide content (lysis samples; n=9 from different passage numbers) or peptide secretion in response to buffer (baseline), and after stimulation with 50 mM KCl or 10 mM glucose + 10 µM forskolin/3-isobutyl-1-methylxanthine (n=6 also different passage numbers). All cell lines produced and processed proglucagon into GLP-1, GLP-2, glicentin and oxyntomodulin in a pattern (prohormone convertase (PC)1/3 dependent) similar to that described for human gut. All three cell lines showed basal secretion of GLP-1 and -2 which increased after stimulation. In contrast to freshly isolated murine L-cells, all lines also expressed PC2 and secreted large amounts of pancreatic glucagon. Neurotensin and somatostatin storage was low and secretion was not consistently increased by stimulation. STC-1 cells released more glucose-dependent insulinotropic polypeptide (GIP) than GLP-1 at baseline (P<0.01) and KCl elevated its secretion (P<0.05). PYY, which normally co-localizes with GLP-1 in distal L-cells, was not detected in any of the cell lines. GLUTag and STC-1 cells also expressed vasoactive intestinal peptide, but none expressed pancreatic polypeptide or insulin. GLUTag contained and secreted large amounts of cholecystokinin while NCI-H716 did not store this peptide and STC-1 contained low amounts. Our results show that hormone production in cell line models of the L-cell have limited similarity to the natural L-cells.

AB - GLUTag, NCI-H716 and STC-1 are cell lines that are widely used to study mechanisms underlying secretion of glucagon-like peptide (GLP-1), but the extent to which they resemble native L-cells is unknown. We used validated immunoassays for 14 different hormones to analyze peptide content (lysis samples; n=9 from different passage numbers) or peptide secretion in response to buffer (baseline), and after stimulation with 50 mM KCl or 10 mM glucose + 10 µM forskolin/3-isobutyl-1-methylxanthine (n=6 also different passage numbers). All cell lines produced and processed proglucagon into GLP-1, GLP-2, glicentin and oxyntomodulin in a pattern (prohormone convertase (PC)1/3 dependent) similar to that described for human gut. All three cell lines showed basal secretion of GLP-1 and -2 which increased after stimulation. In contrast to freshly isolated murine L-cells, all lines also expressed PC2 and secreted large amounts of pancreatic glucagon. Neurotensin and somatostatin storage was low and secretion was not consistently increased by stimulation. STC-1 cells released more glucose-dependent insulinotropic polypeptide (GIP) than GLP-1 at baseline (P<0.01) and KCl elevated its secretion (P<0.05). PYY, which normally co-localizes with GLP-1 in distal L-cells, was not detected in any of the cell lines. GLUTag and STC-1 cells also expressed vasoactive intestinal peptide, but none expressed pancreatic polypeptide or insulin. GLUTag contained and secreted large amounts of cholecystokinin while NCI-H716 did not store this peptide and STC-1 contained low amounts. Our results show that hormone production in cell line models of the L-cell have limited similarity to the natural L-cells.

U2 - 10.1530/JME-15-0293

DO - 10.1530/JME-15-0293

M3 - Journal article

C2 - 26819328

VL - 56

SP - 201

EP - 211

JO - Journal of Molecular Endocrinology

JF - Journal of Molecular Endocrinology

SN - 0952-5041

ER -