Detail
Article
Online article
FT
Medvik - BMC
  • Something wrong with this record ?

Identification of AGR2 Gene-Specific Expression Patterns Associated with Epithelial-Mesenchymal Transition

A. Martisova, L. Sommerova, A. Krejci, I. Selingerova, T. Kolarova, F. Zavadil Kokas, M. Holanek, J. Podhorec, T. Kazda, R. Hrstka

. 2022 ; 23 (18) : . [pub] 20220916

Language English Country Switzerland

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc22024392

Grant support
LX22NPO5102 European Union - Next Generation EU
MMCI, 00209805 Ministry of Health, Czech Republic
Talent Scholarship Brno City Municipality

The TGF-β signaling pathway is involved in numerous cellular processes, and its deregulation may result in cancer development. One of the key processes in tumor progression and metastasis is epithelial to mesenchymal transition (EMT), in which TGF-β signaling plays important roles. Recently, AGR2 was identified as a crucial component of the cellular machinery responsible for maintaining the epithelial phenotype, thereby interfering with the induction of mesenchymal phenotype cells by TGF-β effects in cancer. Here, we performed transcriptomic profiling of A549 lung cancer cells with CRISPR-Cas9 mediated AGR2 knockout with and without TGF-β treatment. We identified significant changes in transcripts associated with focal adhesion and eicosanoid production, in particular arachidonic acid metabolism. Changes in transcripts associated with the focal adhesion pathway were validated by RT-qPCR of COL4A1, COL4A2, FLNA, VAV3, VEGFA, and VINC mRNAs. In addition, immunofluorescence showed the formation of stress fibers and vinculin foci in cells without AGR2 and in response to TGF-β treatment, with synergistic effects observed. These findings imply that both AGR2 downregulation and TGF-β have a role in focal adhesion formation and cancer cell migration and invasion. Transcripts associated with arachidonic acid metabolism were downregulated after both AGR2 knockout and TGF-β treatment and were validated by RT-qPCR of GPX2, PTGS2, and PLA2G4A. Since PGE2 is a product of arachidonic acid metabolism, its lowered concentration in media from AGR2-knockout cells was confirmed by ELISA. Together, our results demonstrate that AGR2 downregulation and TGF-β have an essential role in focal adhesion formation; moreover, we have identified AGR2 as an important component of the arachidonic acid metabolic pathway.

References provided by Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc22024392
003      
CZ-PrNML
005      
20221031100555.0
007      
ta
008      
221017s2022 sz f 000 0|eng||
009      
AR
024    7_
$a 10.3390/ijms231810845 $2 doi
035    __
$a (PubMed)36142758
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a sz
100    1_
$a Martisova, Andrea $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $u National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic $1 https://orcid.org/0000000200810498
245    10
$a Identification of AGR2 Gene-Specific Expression Patterns Associated with Epithelial-Mesenchymal Transition / $c A. Martisova, L. Sommerova, A. Krejci, I. Selingerova, T. Kolarova, F. Zavadil Kokas, M. Holanek, J. Podhorec, T. Kazda, R. Hrstka
520    9_
$a The TGF-β signaling pathway is involved in numerous cellular processes, and its deregulation may result in cancer development. One of the key processes in tumor progression and metastasis is epithelial to mesenchymal transition (EMT), in which TGF-β signaling plays important roles. Recently, AGR2 was identified as a crucial component of the cellular machinery responsible for maintaining the epithelial phenotype, thereby interfering with the induction of mesenchymal phenotype cells by TGF-β effects in cancer. Here, we performed transcriptomic profiling of A549 lung cancer cells with CRISPR-Cas9 mediated AGR2 knockout with and without TGF-β treatment. We identified significant changes in transcripts associated with focal adhesion and eicosanoid production, in particular arachidonic acid metabolism. Changes in transcripts associated with the focal adhesion pathway were validated by RT-qPCR of COL4A1, COL4A2, FLNA, VAV3, VEGFA, and VINC mRNAs. In addition, immunofluorescence showed the formation of stress fibers and vinculin foci in cells without AGR2 and in response to TGF-β treatment, with synergistic effects observed. These findings imply that both AGR2 downregulation and TGF-β have a role in focal adhesion formation and cancer cell migration and invasion. Transcripts associated with arachidonic acid metabolism were downregulated after both AGR2 knockout and TGF-β treatment and were validated by RT-qPCR of GPX2, PTGS2, and PLA2G4A. Since PGE2 is a product of arachidonic acid metabolism, its lowered concentration in media from AGR2-knockout cells was confirmed by ELISA. Together, our results demonstrate that AGR2 downregulation and TGF-β have an essential role in focal adhesion formation; moreover, we have identified AGR2 as an important component of the arachidonic acid metabolic pathway.
650    _2
$a kyselina arachidonová $7 D016718
650    _2
$a nádorové buněčné linie $7 D045744
650    _2
$a pohyb buněk $x genetika $7 D002465
650    _2
$a cyklooxygenasa 2 $x genetika $7 D051546
650    12
$a epitelo-mezenchymální tranzice $x genetika $7 D058750
650    12
$a regulace genové exprese u nádorů $7 D015972
650    _2
$a prostaglandiny E $7 D011458
650    _2
$a transformující růstový faktor beta $x genetika $7 D016212
650    _2
$a vinkulin $x genetika $7 D016596
655    _2
$a časopisecké články $7 D016428
700    1_
$a Sommerova, Lucia $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic
700    1_
$a Krejci, Adam $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic
700    1_
$a Selingerova, Iveta $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $1 https://orcid.org/0000000337133504
700    1_
$a Kolarova, Tamara $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic
700    1_
$a Zavadil Kokas, Filip $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic
700    1_
$a Holanek, Milos $u Department of Comprehensive Cancer Care, Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $u Department of Comprehensive Cancer Care, Faculty of Medicine, Masaryk University, Kamenice 753/5, 62500 Brno, Czech Republic $1 https://orcid.org/0000000312261593
700    1_
$a Podhorec, Jan $u Department of Comprehensive Cancer Care, Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic
700    1_
$a Kazda, Tomas $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $u Department of Radiation Oncology, Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $1 https://orcid.org/0000000172888975
700    1_
$a Hrstka, Roman $u Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty Kopec 7, 65653 Brno, Czech Republic $u Department of Comprehensive Cancer Care, Faculty of Medicine, Masaryk University, Kamenice 753/5, 62500 Brno, Czech Republic $1 https://orcid.org/0000000261392664
773    0_
$w MED00176142 $t International journal of molecular sciences $x 1422-0067 $g Roč. 23, č. 18 (2022)
856    41
$u https://pubmed.ncbi.nlm.nih.gov/36142758 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y p $z 0
990    __
$a 20221017 $b ABA008
991    __
$a 20221031100553 $b ABA008
999    __
$a ok $b bmc $g 1854230 $s 1175682
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2022 $b 23 $c 18 $e 20220916 $i 1422-0067 $m International journal of molecular sciences $n Int J Mol Sci $x MED00176142
GRA    __
$a LX22NPO5102 $p European Union - Next Generation EU
GRA    __
$a MMCI, 00209805 $p Ministry of Health, Czech Republic
GRA    __
$a Talent Scholarship $p Brno City Municipality
LZP    __
$a Pubmed-20221017

Find record

Citation metrics

Archiving options