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Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line

Basic research on types 1 and 2 diabetes mellitus require early stage studies using beta cells or cell lines, ideally of human origin and with preserved insulin secretion in response to glucose. The 1.1E7 cells are a hybrid cell line resulting from the electrofusion of dispersed human islets and PAN...

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Autores principales: Diaz-Ganete, Antonia, Quiroga-de-Castro, Aranzazu, Mateos, Rosa M., Medina, Francisco, Segundo, Carmen, Lechuga-Sancho, Alfonso M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961802/
https://www.ncbi.nlm.nih.gov/pubmed/33806355
http://dx.doi.org/10.3390/ijms22052559
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author Diaz-Ganete, Antonia
Quiroga-de-Castro, Aranzazu
Mateos, Rosa M.
Medina, Francisco
Segundo, Carmen
Lechuga-Sancho, Alfonso M.
author_facet Diaz-Ganete, Antonia
Quiroga-de-Castro, Aranzazu
Mateos, Rosa M.
Medina, Francisco
Segundo, Carmen
Lechuga-Sancho, Alfonso M.
author_sort Diaz-Ganete, Antonia
collection PubMed
description Basic research on types 1 and 2 diabetes mellitus require early stage studies using beta cells or cell lines, ideally of human origin and with preserved insulin secretion in response to glucose. The 1.1E7 cells are a hybrid cell line resulting from the electrofusion of dispersed human islets and PANC-1 cells, capable of secreting insulin in response to glucose, but their survival and function under toxic conditions remains untested. This characterization is the purpose of the present study. We treated these cells with a cytokine mix, high glucose, palmitate, and the latter two combined. Under these conditions, we measured cell viability and apoptosis (MTT, Caspase Glo and TUNEL assays, as well as caspase-8 and -9 levels by Western blotting), endoplasmic reticulum stress markers (EIF2AK3, HSPA4, EIF2a, and HSPA5) by real-time PCR, and insulin secretion with a glucose challenge. All of these stimuli (i) induce apoptosis and ER stress markers expression, (ii) reduce mRNA amounts of 2–5 components of genes involved in the insulin secretory pathway, and (iii) abrogate the insulin release capability of 1.1E7 cells in response to glucose. The most pronounced effects were observed with cytokines and with palmitate and high glucose combined. This characterization may well serve as the starting point for those choosing this cell line for future basic research on certain aspects of diabetes.
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spelling pubmed-79618022021-03-17 Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line Diaz-Ganete, Antonia Quiroga-de-Castro, Aranzazu Mateos, Rosa M. Medina, Francisco Segundo, Carmen Lechuga-Sancho, Alfonso M. Int J Mol Sci Article Basic research on types 1 and 2 diabetes mellitus require early stage studies using beta cells or cell lines, ideally of human origin and with preserved insulin secretion in response to glucose. The 1.1E7 cells are a hybrid cell line resulting from the electrofusion of dispersed human islets and PANC-1 cells, capable of secreting insulin in response to glucose, but their survival and function under toxic conditions remains untested. This characterization is the purpose of the present study. We treated these cells with a cytokine mix, high glucose, palmitate, and the latter two combined. Under these conditions, we measured cell viability and apoptosis (MTT, Caspase Glo and TUNEL assays, as well as caspase-8 and -9 levels by Western blotting), endoplasmic reticulum stress markers (EIF2AK3, HSPA4, EIF2a, and HSPA5) by real-time PCR, and insulin secretion with a glucose challenge. All of these stimuli (i) induce apoptosis and ER stress markers expression, (ii) reduce mRNA amounts of 2–5 components of genes involved in the insulin secretory pathway, and (iii) abrogate the insulin release capability of 1.1E7 cells in response to glucose. The most pronounced effects were observed with cytokines and with palmitate and high glucose combined. This characterization may well serve as the starting point for those choosing this cell line for future basic research on certain aspects of diabetes. MDPI 2021-03-04 /pmc/articles/PMC7961802/ /pubmed/33806355 http://dx.doi.org/10.3390/ijms22052559 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Diaz-Ganete, Antonia
Quiroga-de-Castro, Aranzazu
Mateos, Rosa M.
Medina, Francisco
Segundo, Carmen
Lechuga-Sancho, Alfonso M.
Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title_full Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title_fullStr Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title_full_unstemmed Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title_short Toxicity Induced by Cytokines, Glucose, and Lipids Increase Apoptosis and Hamper Insulin Secretion in the 1.1E7 Beta Cell-Line
title_sort toxicity induced by cytokines, glucose, and lipids increase apoptosis and hamper insulin secretion in the 1.1e7 beta cell-line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961802/
https://www.ncbi.nlm.nih.gov/pubmed/33806355
http://dx.doi.org/10.3390/ijms22052559
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