Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells

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Standard

Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells. / La Barbera, Giorgia; Capriotti, Anna Laura; Michelini, Elisa; Piovesana, Susy; Calabretta, Maria Maddalena; Zenezini Chiozzi, Riccardo; Roda, Aldo; Laganà, Aldo.

I: Proteomics, Bind 17, Nr. 15-16, 1700081, 08.2017.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

La Barbera, G, Capriotti, AL, Michelini, E, Piovesana, S, Calabretta, MM, Zenezini Chiozzi, R, Roda, A & Laganà, A 2017, 'Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells', Proteomics, bind 17, nr. 15-16, 1700081. https://doi.org/10.1002/pmic.201700081

APA

La Barbera, G., Capriotti, A. L., Michelini, E., Piovesana, S., Calabretta, M. M., Zenezini Chiozzi, R., ... Laganà, A. (2017). Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells. Proteomics, 17(15-16), [1700081]. https://doi.org/10.1002/pmic.201700081

Vancouver

La Barbera G, Capriotti AL, Michelini E, Piovesana S, Calabretta MM, Zenezini Chiozzi R o.a. Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells. Proteomics. 2017 aug;17(15-16). 1700081. https://doi.org/10.1002/pmic.201700081

Author

La Barbera, Giorgia ; Capriotti, Anna Laura ; Michelini, Elisa ; Piovesana, Susy ; Calabretta, Maria Maddalena ; Zenezini Chiozzi, Riccardo ; Roda, Aldo ; Laganà, Aldo. / Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells. I: Proteomics. 2017 ; Bind 17, Nr. 15-16.

Bibtex

@article{e9328608b03e4b93a31fc23b97b959e1,
title = "Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells",
abstract = "Microgravity is one of the most important features in spaceflight. Previous evidence from in-vitro studies has shown that significant changes occur under simulated microgravity. For this reason, human colon adenocarcinoma Caco-2 cells were selected as cell model of intestinal epithelial barrier and their response to altered gravity conditions was investigated, especially on the protein level. In this study, we combined label-free shotgun proteomics and bioluminescent reporter gene assays to identify key proteins and pathways involved in the response of Caco-2 cells under reference and microgravity conditions. A two-dimensional clinostat was modified with 3D-printed adaptors to hold conventional T25 culture flasks. The comparative proteome analysis led to identify 38 and 26 proteins differently regulated by simulated microgravity after 48 and 72 h, respectively. Substantial fractions of these proteins are involved in regulation, cellular and metabolic processes and localization. Bioluminescent reporter gene assays were carried out to investigate microgavity-induced alterations on the transcriptional regulation of key targets, such as NF-kB pathway and CYP27A1. While no significant difference was found in the basal transcription, a lower NF-kB basal activation in simulated microgravity conditions was reported, corroborating the hypothesis of reduced immunity in microgravity conditions.",
keywords = "Bioluminescence, Caco-2 cells, Label-free proteomics, Nuclear factor κ-B, Reporter gene technology, Simulated microgravity",
author = "{La Barbera}, Giorgia and Capriotti, {Anna Laura} and Elisa Michelini and Susy Piovesana and Calabretta, {Maria Maddalena} and {Zenezini Chiozzi}, Riccardo and Aldo Roda and Aldo Lagan{\`a}",
year = "2017",
month = "8",
doi = "10.1002/pmic.201700081",
language = "English",
volume = "17",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",
number = "15-16",

}

RIS

TY - JOUR

T1 - Proteomic analysis and bioluminescent reporter gene assays to investigate effects of simulated microgravity on Caco-2 cells

AU - La Barbera, Giorgia

AU - Capriotti, Anna Laura

AU - Michelini, Elisa

AU - Piovesana, Susy

AU - Calabretta, Maria Maddalena

AU - Zenezini Chiozzi, Riccardo

AU - Roda, Aldo

AU - Laganà, Aldo

PY - 2017/8

Y1 - 2017/8

N2 - Microgravity is one of the most important features in spaceflight. Previous evidence from in-vitro studies has shown that significant changes occur under simulated microgravity. For this reason, human colon adenocarcinoma Caco-2 cells were selected as cell model of intestinal epithelial barrier and their response to altered gravity conditions was investigated, especially on the protein level. In this study, we combined label-free shotgun proteomics and bioluminescent reporter gene assays to identify key proteins and pathways involved in the response of Caco-2 cells under reference and microgravity conditions. A two-dimensional clinostat was modified with 3D-printed adaptors to hold conventional T25 culture flasks. The comparative proteome analysis led to identify 38 and 26 proteins differently regulated by simulated microgravity after 48 and 72 h, respectively. Substantial fractions of these proteins are involved in regulation, cellular and metabolic processes and localization. Bioluminescent reporter gene assays were carried out to investigate microgavity-induced alterations on the transcriptional regulation of key targets, such as NF-kB pathway and CYP27A1. While no significant difference was found in the basal transcription, a lower NF-kB basal activation in simulated microgravity conditions was reported, corroborating the hypothesis of reduced immunity in microgravity conditions.

AB - Microgravity is one of the most important features in spaceflight. Previous evidence from in-vitro studies has shown that significant changes occur under simulated microgravity. For this reason, human colon adenocarcinoma Caco-2 cells were selected as cell model of intestinal epithelial barrier and their response to altered gravity conditions was investigated, especially on the protein level. In this study, we combined label-free shotgun proteomics and bioluminescent reporter gene assays to identify key proteins and pathways involved in the response of Caco-2 cells under reference and microgravity conditions. A two-dimensional clinostat was modified with 3D-printed adaptors to hold conventional T25 culture flasks. The comparative proteome analysis led to identify 38 and 26 proteins differently regulated by simulated microgravity after 48 and 72 h, respectively. Substantial fractions of these proteins are involved in regulation, cellular and metabolic processes and localization. Bioluminescent reporter gene assays were carried out to investigate microgavity-induced alterations on the transcriptional regulation of key targets, such as NF-kB pathway and CYP27A1. While no significant difference was found in the basal transcription, a lower NF-kB basal activation in simulated microgravity conditions was reported, corroborating the hypothesis of reduced immunity in microgravity conditions.

KW - Bioluminescence

KW - Caco-2 cells

KW - Label-free proteomics

KW - Nuclear factor κ-B

KW - Reporter gene technology

KW - Simulated microgravity

UR - http://www.scopus.com/inward/record.url?scp=85027859865&partnerID=8YFLogxK

U2 - 10.1002/pmic.201700081

DO - 10.1002/pmic.201700081

M3 - Journal article

C2 - 28727291

AN - SCOPUS:85027859865

VL - 17

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 15-16

M1 - 1700081

ER -

ID: 231311009