In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line.
Type 1 diabetes mellitus (T1DM) is caused by the selective destruction of insulin-producing β-cells. This process is mediated by cells of the immune system through release of nitric oxide, free radicals and pro-inflammatory cytokines, which induce a complex network of intracellular signalling cascad...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3290556?pdf=render |
| _version_ | 1811289891890266112 |
|---|---|
| author | Antonina Coppola Laura Tomasello Giuseppe Pizzolanti Ida Pucci-Minafra Nadia Albanese Gianluca Di Cara Patrizia Cancemi Maria Pitrone Alessandra Bommarito Elvira Carissimi Giovanni Zito Angela Criscimanna Aldo Galluzzo Carla Giordano |
| author_facet | Antonina Coppola Laura Tomasello Giuseppe Pizzolanti Ida Pucci-Minafra Nadia Albanese Gianluca Di Cara Patrizia Cancemi Maria Pitrone Alessandra Bommarito Elvira Carissimi Giovanni Zito Angela Criscimanna Aldo Galluzzo Carla Giordano |
| author_sort | Antonina Coppola |
| collection | DOAJ |
| description | Type 1 diabetes mellitus (T1DM) is caused by the selective destruction of insulin-producing β-cells. This process is mediated by cells of the immune system through release of nitric oxide, free radicals and pro-inflammatory cytokines, which induce a complex network of intracellular signalling cascades, eventually affecting the expression of genes involved in β-cell survival.The aim of our study was to investigate possible mechanisms of resistance to cytokine-induced β-cell death. To this purpose, we created a cytokine-resistant β-cell line (β-TC3R) by chronically treating the β-TC3 murine insulinoma cell line with IL-1β + IFN-γ. β-TC3R cells exhibited higher proliferation rate and resistance to cytokine-mediated cell death in comparison to the parental line. Interestingly, they maintained expression of β-cell specific markers, such as PDX1, NKX6.1, GLUT2 and insulin. The analysis of the secretory function showed that β-TC3R cells have impaired glucose-induced c-peptide release, which however was only moderately reduced after incubation with KCl and tolbutamide. Gene expression analysis showed that β-TC3R cells were characterized by downregulation of IL-1β and IFN-γ receptors and upregulation of SOCS3, the classical negative regulator of cytokines signaling. Comparative proteomic analysis showed specific upregulation of 35 proteins, mainly involved in cell death, stress response and folding. Among them, SUMO4, a negative feedback regulator in NF-kB and JAK/STAT signaling pathways, resulted hyper-expressed. Silencing of SUMO4 was able to restore sensitivity to cytokine-induced cell death in β-TC3R cells, suggesting it may play a key role in acquired cytokine resistance by blocking JAK/STAT and NF-kB lethal signaling.In conclusion, our study represents the first extensive proteomic characterization of a murine cytokine-resistant β-cell line, which might represent a useful tool for studying the mechanisms involved in resistance to cytokine-mediated β-cell death. This knowledge may be of potential benefit for patients with T1DM. In particular, SUMO4 could be used as a therapeutical target. |
| first_indexed | 2024-04-13T04:03:15Z |
| format | Article |
| id | doaj.art-5e78d116e77c45cf8b9505f9ebb17946 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T04:03:15Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-5e78d116e77c45cf8b9505f9ebb179462022-12-22T03:03:25ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0172e3210910.1371/journal.pone.0032109In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line.Antonina CoppolaLaura TomaselloGiuseppe PizzolantiIda Pucci-MinafraNadia AlbaneseGianluca Di CaraPatrizia CancemiMaria PitroneAlessandra BommaritoElvira CarissimiGiovanni ZitoAngela CriscimannaAldo GalluzzoCarla GiordanoType 1 diabetes mellitus (T1DM) is caused by the selective destruction of insulin-producing β-cells. This process is mediated by cells of the immune system through release of nitric oxide, free radicals and pro-inflammatory cytokines, which induce a complex network of intracellular signalling cascades, eventually affecting the expression of genes involved in β-cell survival.The aim of our study was to investigate possible mechanisms of resistance to cytokine-induced β-cell death. To this purpose, we created a cytokine-resistant β-cell line (β-TC3R) by chronically treating the β-TC3 murine insulinoma cell line with IL-1β + IFN-γ. β-TC3R cells exhibited higher proliferation rate and resistance to cytokine-mediated cell death in comparison to the parental line. Interestingly, they maintained expression of β-cell specific markers, such as PDX1, NKX6.1, GLUT2 and insulin. The analysis of the secretory function showed that β-TC3R cells have impaired glucose-induced c-peptide release, which however was only moderately reduced after incubation with KCl and tolbutamide. Gene expression analysis showed that β-TC3R cells were characterized by downregulation of IL-1β and IFN-γ receptors and upregulation of SOCS3, the classical negative regulator of cytokines signaling. Comparative proteomic analysis showed specific upregulation of 35 proteins, mainly involved in cell death, stress response and folding. Among them, SUMO4, a negative feedback regulator in NF-kB and JAK/STAT signaling pathways, resulted hyper-expressed. Silencing of SUMO4 was able to restore sensitivity to cytokine-induced cell death in β-TC3R cells, suggesting it may play a key role in acquired cytokine resistance by blocking JAK/STAT and NF-kB lethal signaling.In conclusion, our study represents the first extensive proteomic characterization of a murine cytokine-resistant β-cell line, which might represent a useful tool for studying the mechanisms involved in resistance to cytokine-mediated β-cell death. This knowledge may be of potential benefit for patients with T1DM. In particular, SUMO4 could be used as a therapeutical target.http://europepmc.org/articles/PMC3290556?pdf=render |
| spellingShingle | Antonina Coppola Laura Tomasello Giuseppe Pizzolanti Ida Pucci-Minafra Nadia Albanese Gianluca Di Cara Patrizia Cancemi Maria Pitrone Alessandra Bommarito Elvira Carissimi Giovanni Zito Angela Criscimanna Aldo Galluzzo Carla Giordano In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. PLoS ONE |
| title | In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. |
| title_full | In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. |
| title_fullStr | In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. |
| title_full_unstemmed | In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. |
| title_short | In vitro phenotypic, genomic and proteomic characterization of a cytokine-resistant murine β-TC3 cell line. |
| title_sort | in vitro phenotypic genomic and proteomic characterization of a cytokine resistant murine β tc3 cell line |
| url | http://europepmc.org/articles/PMC3290556?pdf=render |
| work_keys_str_mv | AT antoninacoppola invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT lauratomasello invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT giuseppepizzolanti invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT idapucciminafra invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT nadiaalbanese invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT gianlucadicara invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT patriziacancemi invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT mariapitrone invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT alessandrabommarito invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT elviracarissimi invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT giovannizito invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT angelacriscimanna invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT aldogalluzzo invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline AT carlagiordano invitrophenotypicgenomicandproteomiccharacterizationofacytokineresistantmurinebtc3cellline |