Desmocollin-1 is associated with pro-metastatic phenotype of
luminal A breast cancer cells and is modulated by parthenolide

a 2024

Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide

LAPČÍK, Petr; Petr ŠULC; Lucia JANÁČOVÁ; Kateřina JÍLKOVÁ; David POTĚŠIL et al.

Základní údaje

Originální název

Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide

Autoři

LAPČÍK, Petr; Petr ŠULC; Lucia JANÁČOVÁ; Kateřina JÍLKOVÁ; David POTĚŠIL; Pavla BOUCHALOVÁ; Petr MÜLLER a Pavel BOUCHAL

Vydání

In Book of Abstracts of 10th Informal Proteomic Meeting, Olomouc 28.-29.11.2024, Josef Chmelik Prize, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Označené pro přenos do RIV

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

DIA, Proteomics, Pull-down, DSC1, Breast cancer, Metastasis

Příznaky

Mezinárodní význam

Změněno: 5. 12. 2024 11:40, Mgr. Petr Lapčík, Ph.D.

Anotace

V originále

Background: Desmocollin-1 (DSC1) is a desmosomal transmembrane glycoprotein that maintains cell-to-cell adhesion. DSC1 was previously associated with lymph node metastasis of luminal A breast tumors and was found to increase migration and invasion of MCF7 cells in vitro. Therefore, we focused on DSC1 role in cellular and molecular mechanisms in luminal A breast cancer and its possible therapeutic modulation. Methods: Western blotting was used to select potential inhibitor decreasing DSC1 protein level in MCF7 cell line. Using atomic force microscopy we evaluated effect of DSC1 overexpression and modulation on cell morphology. The LC-MS/MS analysis of total proteome on Orbitrap Lumos and RNA-Seq analysis of total transcriptome on Illumina NextSeq 500 were performed to study the molecular mechanisms associated with DSC1. Pull-down analysis with LC-MS/MS detection was carried out to uncover DSC1 protein interactome in MCF7 cells. Results: Analysis of DSC1 protein levels in response to selected inhibitors displays significant DSC1 downregulation (p-value ≤ 0.01) in MCF7 cells treated with NF-κB inhibitor parthenolide. Analysis of mechanic cell properties in response to DSC1 overexpression and parthenolide treatment using atomic force microscopy reveals that DSC1 overexpression reduces height of MCF7 cells and conversely, parthenolide decreases cell stiffness of MCF7 cells overexpressing DSC1. The LC-MS/MS total proteome analysis in data-independent acquisition mode shows a strong connection between DSC1 overexpression and increased levels of proteins LACRT and IGFBP5, increased expression of IGFBP5 is confirmed by RNA-Seq. Pathway analysis of proteomics data uncovers enrichment of proliferative MCM_BIOCARTA pathway including CDK2 and MCM2-7 after DSC1 overexpression. Parthenolide decreases expression of LACRT, IGFBP5 and MCM_BIOCARTA pathway specifically in DSC1 overexpressing cells. Pull-down assay identifies DSC1 interactions with cadherin family proteins including DSG2, CDH1, CDH3 and tyrosine kinase receptors HER2 and HER3; parthenolide modulates DSC1-HER3 interaction. Conclusions: Our systems biology data indicate that DSC1 is connected to mechanisms of cell cycle regulation in luminal A breast cancer cells, and can be effectively modulated by parthenolide. This work was supported by Ministry of Health of the Czech Republic, grant nr. NU22-08-00230, all rights reserved. CIISB, Instruct-CZ Centre of Instruct-ERIC EU consortium, funded by MEYS CR infrastructure project LM2023042 and European Regional Development Fund-Project „UP CIISB “ (No. CZ.02.1.01/0.0/0.0/18_046/0015974), is gratefully acknowledged for the financial support of the measurements at the CEITEC Proteomics Core Facility and for the AFM measurements at Nanobiotechnology core facility. Bioinformatics Core Facility of CEITEC Masaryk University supported by the NCMG research infrastructure (LM2018132 funded by MEYS CR) is acknowledged for RNA-Seq data analysis. Supported by the project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102)—Funded by the European Union—Next Generation EU.

Návaznosti

LM2018132, projekt VaV

Název: Národní centrum lékařské genomiky (Akronym: NCLG)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Národní centrum lékařské genomiky

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

LX22NPO5102, projekt VaV

Název: Národní ústav pro výzkum rakoviny (Akronym: NÚVR)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Národní ústav pro výzkum rakoviny, 5.1 EXCELES

NU22-08-00230, projekt VaV

Název: Proteogenomová klasifikace trojitě negativních nádorů prsu ve vztahu k prognóze a cílené terapii

Investor: Ministerstvo zdravotnictví ČR, Proteogenomová klasifikace trojitě negativních nádorů prsu ve vztahu k prognóze a cílené terapii, Podprogram 1 - standardní

Citovat

LAPČÍK, Petr; Petr ŠULC; Lucia JANÁČOVÁ; Kateřina JÍLKOVÁ; David POTĚŠIL; Pavla BOUCHALOVÁ; Petr MÜLLER a Pavel BOUCHAL. Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide. In In Book of Abstracts of 10th Informal Proteomic Meeting, Olomouc 28.-29.11.2024, Josef Chmelik Prize. 2024.

@proceedings{2457540,
   author = {Lapčík, Petr and Šulc, Petr and Janáčová, Lucia and Jílková, Kateřina and Potěšil, David and Bouchalová, Pavla and Müller, Petr and Bouchal, Pavel},
   booktitle = {In Book of Abstracts of 10th Informal Proteomic Meeting, Olomouc 28.-29.11.2024, Josef Chmelik Prize},
   keywords = {DIA, Proteomics, Pull-down, DSC1, Breast cancer, Metastasis},
   language = {eng},
   title = {Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide},
   url = {https://drive.google.com/file/d/1Ful5TBEr6q22Nn86JWEklTh9ep21X2zZ/view},
   year = {2024}
}

TY  - CONF
ID  - 2457540
AU  - Lapčík, Petr - Šulc, Petr - Janáčová, Lucia - Jílková, Kateřina - Potěšil, David - Bouchalová, Pavla - Müller, Petr - Bouchal, Pavel
PY  - 2024
TI  - Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide
KW  - DIA, Proteomics, Pull-down, DSC1, Breast cancer, Metastasis
UR  - https://drive.google.com/file/d/1Ful5TBEr6q22Nn86JWEklTh9ep21X2zZ/view
N2  - Background: Desmocollin-1 (DSC1) is a desmosomal transmembrane glycoprotein that maintains cell-to-cell adhesion. DSC1 was previously associated with lymph node metastasis of luminal A breast tumors and was found to increase migration and invasion of MCF7 cells in vitro. Therefore, we focused on DSC1 role in cellular and molecular mechanisms in luminal A breast cancer and its possible therapeutic modulation. Methods: Western blotting was used to select potential inhibitor decreasing DSC1 protein level in MCF7 cell line. Using atomic force microscopy we evaluated effect of DSC1 overexpression and modulation on cell morphology. The LC-MS/MS analysis of total proteome on Orbitrap Lumos and RNA-Seq analysis of total transcriptome on Illumina NextSeq 500 were performed to study the molecular mechanisms associated with DSC1. Pull-down analysis with LC-MS/MS detection was carried out to uncover DSC1 protein interactome in MCF7 cells. Results: Analysis of DSC1 protein levels in response to selected inhibitors displays significant DSC1 downregulation (p-value ≤ 0.01) in MCF7 cells treated with NF-κB inhibitor parthenolide. Analysis of mechanic cell properties in response to DSC1 overexpression and parthenolide treatment using atomic force microscopy reveals that DSC1 overexpression reduces height of MCF7 cells and conversely, parthenolide decreases cell stiffness of MCF7 cells overexpressing DSC1. The LC-MS/MS total proteome analysis in data-independent acquisition mode shows a strong connection between DSC1 overexpression and increased levels of proteins LACRT and IGFBP5, increased expression of IGFBP5 is confirmed by RNA-Seq. Pathway analysis of proteomics data uncovers enrichment of proliferative MCM_BIOCARTA pathway including CDK2 and MCM2-7 after DSC1 overexpression. Parthenolide decreases expression of LACRT, IGFBP5 and MCM_BIOCARTA pathway specifically in DSC1 overexpressing cells. Pull-down assay identifies DSC1 interactions with cadherin family proteins including DSG2, CDH1, CDH3 and tyrosine kinase receptors HER2 and HER3; parthenolide modulates DSC1-HER3 interaction. Conclusions: Our systems biology data indicate that DSC1 is connected to mechanisms of cell cycle regulation in luminal A breast cancer cells, and can be effectively modulated by parthenolide. This work was supported by Ministry of Health of the Czech Republic, grant nr. NU22-08-00230, all rights reserved. CIISB, Instruct-CZ Centre of Instruct-ERIC EU consortium, funded by MEYS CR infrastructure project LM2023042 and European Regional Development Fund-Project „UP CIISB “ (No. CZ.02.1.01/0.0/0.0/18_046/0015974), is gratefully acknowledged for the financial support of the measurements at the CEITEC Proteomics Core Facility and for the AFM measurements at Nanobiotechnology core facility. Bioinformatics Core Facility of CEITEC Masaryk University supported by the NCMG research infrastructure (LM2018132 funded by MEYS CR) is acknowledged for RNA-Seq data analysis. Supported by the project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102)—Funded by the European Union—Next Generation EU.
ER  -

LAPČÍK, Petr; Petr ŠULC; Lucia JANÁČOVÁ; Kateřina JÍLKOVÁ; David POTĚŠIL; Pavla BOUCHALOVÁ; Petr MÜLLER a Pavel BOUCHAL. Desmocollin-1 is associated with pro-metastatic phenotype of luminal A breast cancer cells and is modulated by parthenolide. In \textit{In Book of Abstracts of 10th Informal Proteomic Meeting, Olomouc 28.-29.11.2024, Josef Chmelik Prize}. 2024.

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