Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model

Faragó, Anna; Zvara, Ágnes; Tiszlavicz, László; Hunyadi-Gulyás, Éva; Darula, Zsuzsanna; Hegedűs, Zoltán; Szabó, Enikő; Surguta, Sára Eszter; Tóvári, József; Puskás, László G.; Szebeni, Gábor J.

doi:10.3390/ijms25074022

Open AccessArticle

Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model

by

Anna Faragó

^1,2,

Ágnes Zvara

^3,4,

László Tiszlavicz

⁵,

Éva Hunyadi-Gulyás

^4,6

,

Zsuzsanna Darula

^4,6,7

,

Zoltán Hegedűs

^4,8,9

,

Enikő Szabó

^3,4,

Sára Eszter Surguta

¹⁰,

József Tóvári

¹⁰,

László G. Puskás

^{3,4,11,12,*,†} and

Gábor J. Szebeni

^{1,3,4,13,*,†}

¹

Astridbio Technologies Ltd., Wimmer Fülöp utca 1, H6728 Szeged, Hungary

²

University of Szeged, Albert Szent-Györgyi Medical School, Doctoral School of Multidisciplinary Medical Sciences, Dóm tér 9, H6720 Szeged, Hungary

³

Institute of Genetics, Laboratory of Functional Genomics, HUN-REN Biological Research Centre, Temesvári krt. 62, H6726 Szeged, Hungary

⁴

Core Facility HUN-REN Biological Research Centre, Temesvári krt. 62, H6726 Szeged, Hungary

⁵

Department of Pathology, University of Szeged, Állomás u. 2, H6725 Szeged, Hungary

⁶

Laboratory of Proteomics Research, HUN-REN Biological Research Centre, Temesvári krt. 62, H6726 Szeged, Hungary

⁷

The Hungarian Centre of Excellence for Molecular Medicine (HCEMM) Single Cell Omics Advanced Core Facility, Biological Research Centre, Temesvári krt. 62, H6726 Szeged, Hungary

⁸

Laboratory of Bioinformatics, HUN-REN Biological Research Centre, Temesvári krt. 62, H6726 Szeged, Hungary

⁹

Department of Biochemistry and Medical Chemistry, Medical School, University of Pécs, Szigeti út 12, H7624 Pécs, Hungary

¹⁰

Department of Experimental Pharmacology, The National Tumor Biology Laboratory, National Institute of Oncology, Ráth György u. 7-9, H1122 Budapest, Hungary

¹¹

Avidin Ltd., Alsó Kikötő sor 11/D, H6726 Szeged, Hungary

¹²

Avicor Ltd., Alsó Kikötő sor 11/D, H6726 Szeged, Hungary

¹³

Department of Internal Medicine, Hematology Centre, Faculty of Medicine University of Szeged, H6725 Szeged, Hungary

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^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2024, 25(7), 4022; https://doi.org/10.3390/ijms25074022

Submission received: 23 February 2024 / Revised: 27 March 2024 / Accepted: 3 April 2024 / Published: 4 April 2024

(This article belongs to the Special Issue Immunophenotyping in Autoimmune Diseases and Cancer, 3rd Edition)

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Abstract

A murine colorectal carcinoma (CRC) model was established. CT26 colon carcinoma cells were injected into BALB/c mice’s spleen to study the primary tumor and the mechanisms of cell spread of colon cancer to the liver. The CRC was verified by the immunohistochemistry of Pan Cytokeratin and Vimentin expression. Immunophenotyping of leukocytes isolated from CRC-bearing BALB/c mice or healthy controls, such as CD19+ B cells, CD11+ myeloid cells, and CD3+ T cells, was carried out using fluorochrome-labeled lectins. The binding of six lectins to white blood cells, such as galectin-1 (Gal1), siglec-1 (Sig1), Sambucus nigra lectin (SNA), Aleuria aurantia lectin (AAL), Phytolacca americana lectin (PWM), and galectin-3 (Gal3), was assayed. Flow cytometric analysis of the splenocytes revealed the increased binding of SNA, and AAL to CD3 + T cells and CD11b myeloid cells; and increased siglec-1 and AAL binding to CD19 B cells of the tumor-bearing mice. The whole proteomic analysis of the established CRC-bearing liver and spleen versus healthy tissues identified differentially expressed proteins, characteristic of the primary or secondary CRC tissues. KEGG Gene Ontology bioinformatic analysis delineated the established murine CRC characteristic protein interaction networks, biological pathways, and cellular processes involved in CRC. Galectin-1 and S100A4 were identified as upregulated proteins in the primary and secondary CT26 tumor tissues, and these were previously reported to contribute to the poor prognosis of CRC patients. Modelling the development of liver colonization of CRC by the injection of CT26 cells into the spleen may facilitate the understanding of carcinogenesis in human CRC and contribute to the development of novel therapeutic strategies.

Keywords: colorectal carcinoma; lectin binding sugar code; proteomic analysis of murine CRC

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MDPI and ACS Style

Faragó, A.; Zvara, Á.; Tiszlavicz, L.; Hunyadi-Gulyás, É.; Darula, Z.; Hegedűs, Z.; Szabó, E.; Surguta, S.E.; Tóvári, J.; Puskás, L.G.; et al. Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model. Int. J. Mol. Sci. 2024, 25, 4022. https://doi.org/10.3390/ijms25074022

AMA Style

Faragó A, Zvara Á, Tiszlavicz L, Hunyadi-Gulyás É, Darula Z, Hegedűs Z, Szabó E, Surguta SE, Tóvári J, Puskás LG, et al. Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model. International Journal of Molecular Sciences. 2024; 25(7):4022. https://doi.org/10.3390/ijms25074022

Chicago/Turabian Style

Faragó, Anna, Ágnes Zvara, László Tiszlavicz, Éva Hunyadi-Gulyás, Zsuzsanna Darula, Zoltán Hegedűs, Enikő Szabó, Sára Eszter Surguta, József Tóvári, László G. Puskás, and et al. 2024. "Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model" International Journal of Molecular Sciences 25, no. 7: 4022. https://doi.org/10.3390/ijms25074022

APA Style

Faragó, A., Zvara, Á., Tiszlavicz, L., Hunyadi-Gulyás, É., Darula, Z., Hegedűs, Z., Szabó, E., Surguta, S. E., Tóvári, J., Puskás, L. G., & Szebeni, G. J. (2024). Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model. International Journal of Molecular Sciences, 25(7), 4022. https://doi.org/10.3390/ijms25074022

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Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model

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