Cargando…

Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal

Claudin-2 (CLDN2), a tight junctional protein, is involved in the chemoresistance in a three-dimensional spheroid culture model of human lung adenocarcinoma A549 cells. However, the mechanism has not been fully clarified. We found that the knockdown of CLDN2 expression by siRNA in the spheroid reduc...

Descripción completa

Detalles Bibliográficos
Autores principales: Ito, Ayaka, Nasako, Haruka, Akizuki, Risa, Takashina, Yui, Eguchi, Hiroaki, Matsunaga, Toshiyuki, Yoshino, Yuta, Endo, Satoshi, Ikari, Akira
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235168/
https://www.ncbi.nlm.nih.gov/pubmed/34205320
http://dx.doi.org/10.3390/ijms22126582
_version_ 1783714253485637632
author Ito, Ayaka
Nasako, Haruka
Akizuki, Risa
Takashina, Yui
Eguchi, Hiroaki
Matsunaga, Toshiyuki
Yoshino, Yuta
Endo, Satoshi
Ikari, Akira
author_facet Ito, Ayaka
Nasako, Haruka
Akizuki, Risa
Takashina, Yui
Eguchi, Hiroaki
Matsunaga, Toshiyuki
Yoshino, Yuta
Endo, Satoshi
Ikari, Akira
author_sort Ito, Ayaka
collection PubMed
description Claudin-2 (CLDN2), a tight junctional protein, is involved in the chemoresistance in a three-dimensional spheroid culture model of human lung adenocarcinoma A549 cells. However, the mechanism has not been fully clarified. We found that the knockdown of CLDN2 expression by siRNA in the spheroid reduces the expression of glucose transporters and metabolic enzymes. In a two-dimensional culture model, the expression of these proteins was increased by glucose deprivation or fasentin, an inhibitor of glucose transporter. In addition, the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and antioxidant enzymes including heme oxygenase-1, NAD(P)H:quinone oxidoreductase-1, and a glutamate–cysteine ligase modifier subunit were increased by fasentin. The fluorescence intensities of JC-1, a probe of mitochondrial membrane potential, and MitoROS 580, a probe of mitochondrial superoxide production, were increased by fasentin. These results suggest that mitochondrial production of reactive oxygen species is increased by glucose deficiency. The knockdown of CLDN2 enhanced the flux of 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG), a fluorescent deoxyglucose derivative, in a transwell assay, and the accumulation of glucose and 2-NBDG in spheroid cells. The expression of Nrf2 was decreased by CLDN2 knockdown, which was inhibited by fasentin and sulforaphane, a typical Nrf2 activator, in spheroid cells. The sensitivity of spheroid cells to doxorubicin, an anthracycline antitumor antibiotic, was enhanced by CLDN2 knockdown, which was inhibited by fasentin and sulforaphane. We suggest that CLDN2 induces chemoresistance in spheroid cells mediated through the inhibition of glucose transport and activation of the Nrf2 signal.
format Online
Article
Text
id pubmed-8235168
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-82351682021-06-27 Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal Ito, Ayaka Nasako, Haruka Akizuki, Risa Takashina, Yui Eguchi, Hiroaki Matsunaga, Toshiyuki Yoshino, Yuta Endo, Satoshi Ikari, Akira Int J Mol Sci Article Claudin-2 (CLDN2), a tight junctional protein, is involved in the chemoresistance in a three-dimensional spheroid culture model of human lung adenocarcinoma A549 cells. However, the mechanism has not been fully clarified. We found that the knockdown of CLDN2 expression by siRNA in the spheroid reduces the expression of glucose transporters and metabolic enzymes. In a two-dimensional culture model, the expression of these proteins was increased by glucose deprivation or fasentin, an inhibitor of glucose transporter. In addition, the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and antioxidant enzymes including heme oxygenase-1, NAD(P)H:quinone oxidoreductase-1, and a glutamate–cysteine ligase modifier subunit were increased by fasentin. The fluorescence intensities of JC-1, a probe of mitochondrial membrane potential, and MitoROS 580, a probe of mitochondrial superoxide production, were increased by fasentin. These results suggest that mitochondrial production of reactive oxygen species is increased by glucose deficiency. The knockdown of CLDN2 enhanced the flux of 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG), a fluorescent deoxyglucose derivative, in a transwell assay, and the accumulation of glucose and 2-NBDG in spheroid cells. The expression of Nrf2 was decreased by CLDN2 knockdown, which was inhibited by fasentin and sulforaphane, a typical Nrf2 activator, in spheroid cells. The sensitivity of spheroid cells to doxorubicin, an anthracycline antitumor antibiotic, was enhanced by CLDN2 knockdown, which was inhibited by fasentin and sulforaphane. We suggest that CLDN2 induces chemoresistance in spheroid cells mediated through the inhibition of glucose transport and activation of the Nrf2 signal. MDPI 2021-06-19 /pmc/articles/PMC8235168/ /pubmed/34205320 http://dx.doi.org/10.3390/ijms22126582 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ito, Ayaka
Nasako, Haruka
Akizuki, Risa
Takashina, Yui
Eguchi, Hiroaki
Matsunaga, Toshiyuki
Yoshino, Yuta
Endo, Satoshi
Ikari, Akira
Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title_full Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title_fullStr Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title_full_unstemmed Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title_short Elevation of Chemosensitivity of Lung Adenocarcinoma A549 Spheroid Cells by Claudin-2 Knockdown through Activation of Glucose Transport and Inhibition of Nrf2 Signal
title_sort elevation of chemosensitivity of lung adenocarcinoma a549 spheroid cells by claudin-2 knockdown through activation of glucose transport and inhibition of nrf2 signal
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235168/
https://www.ncbi.nlm.nih.gov/pubmed/34205320
http://dx.doi.org/10.3390/ijms22126582
work_keys_str_mv AT itoayaka elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT nasakoharuka elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT akizukirisa elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT takashinayui elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT eguchihiroaki elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT matsunagatoshiyuki elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT yoshinoyuta elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT endosatoshi elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal
AT ikariakira elevationofchemosensitivityoflungadenocarcinomaa549spheroidcellsbyclaudin2knockdownthroughactivationofglucosetransportandinhibitionofnrf2signal