دورية أكاديمية
أعرض في EDS

In-depth quantitative proteomics analysis revealed C1GALT1 depletion in ECC-1 cells mimics an aggressive endometrial cancer phenotype observed in cancer patients with low C1GALT1 expression.

التفاصيل البيبلوغرافية
العنوان: In-depth quantitative proteomics analysis revealed C1GALT1 depletion in ECC-1 cells mimics an aggressive endometrial cancer phenotype observed in cancer patients with low C1GALT1 expression.
المؤلفون: Montero-Calle A; Chronic Disease Programme, UFIEC, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain., López-Janeiro Á; Department of Pathology, Hospital Universitario La Paz, 28046, Madrid, Spain., Mendes ML; Department of Infection and Immunity, Luxembourg Institute of Health, 1445, Strassen, Luxembourg., Perez-Hernandez D; Department of Infection and Immunity, Luxembourg Institute of Health, 1445, Strassen, Luxembourg., Echevarría I; Chronic Disease Programme, UFIEC, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain.; Biochemistry and Molecular Biology Department, Facultad de Óptica y Optometría, Universidad Complutense de Madrid, 28037, Madrid, Spain., Ruz-Caracuel I; Department of Pathology, Hospital Universitario La Paz, 28046, Madrid, Spain., Heredia-Soto V; Translational Oncology, La Paz University Hospital (IdiPAZ), 28046, Madrid, Spain.; Center for Biomedical Research in the Cancer Network (Centro de Investigación Biomédica en Red de Cáncer, CIBERONC), Instituto de Salud Carlos III, 28046, Madrid, Spain., Mendiola M; Center for Biomedical Research in the Cancer Network (Centro de Investigación Biomédica en Red de Cáncer, CIBERONC), Instituto de Salud Carlos III, 28046, Madrid, Spain.; Molecular Pathology and Therapeutic Targets Group, La Paz University Hospital (IdiPAZ), 28046, Madrid, Spain., Hardisson D; Department of Pathology, Hospital Universitario La Paz, 28046, Madrid, Spain.; Center for Biomedical Research in the Cancer Network (Centro de Investigación Biomédica en Red de Cáncer, CIBERONC), Instituto de Salud Carlos III, 28046, Madrid, Spain.; Molecular Pathology and Therapeutic Targets Group, La Paz University Hospital (IdiPAZ), 28046, Madrid, Spain.; Faculty of Medicine, Universidad Autónoma de Madrid, 28029, Madrid, Spain., Argüeso P; Tufts Medical Center, Tufts University School of Medicine, Boston, Massachusetts, USA., Peláez-García A; Molecular Pathology and Therapeutic Targets Group, La Paz University Hospital (IdiPAZ), 28046, Madrid, Spain. alberto.pelaez@idipaz.es., Guzman-Aranguez A; Biochemistry and Molecular Biology Department, Facultad de Óptica y Optometría, Universidad Complutense de Madrid, 28037, Madrid, Spain. aguzman@opt.ucm.es., Barderas R; Chronic Disease Programme, UFIEC, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain. r.barderasm@isciii.es.; Functional Proteomics Unit, UFIEC, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain. r.barderasm@isciii.es.
المصدر: Cellular oncology (Dordrecht) [Cell Oncol (Dordr)] 2023 Jun; Vol. 46 (3), pp. 697-715. Date of Electronic Publication: 2023 Feb 06.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer Country of Publication: Netherlands NLM ID: 101552938 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2211-3436 (Electronic) Linking ISSN: 22113428 NLM ISO Abbreviation: Cell Oncol (Dordr) Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Dordrecht : Springer
مواضيع طبية MeSH: Proteomics* , Endometrial Neoplasms*, Humans ; Female ; Glycosylation ; Chromatography, Liquid ; Tandem Mass Spectrometry ; Phenotype ; Galactosyltransferases
مستخلص: Background: Endometrial cancer (EC) is the most common cancer of the female reproductive organs. Despite the good overall prognosis of most low-grade ECs, FIGO I and FIGO II patients might experience tumor recurrence and worse prognosis. The study of alterations related to EC pathogenesis might help to get insights into underlying mechanisms involved in EC development and progression.
Methods: Core tumoral samples were used to investigate the role of C1GALT1 in EC by immunohistochemistry (IHC). ECC-1 cells were used as endometrioid EC model to investigate the effect of C1GALT1 depletion using C1GALT1 specific shRNAs. SILAC quantitative proteomics analyses and cell-based assays, PCR, qPCR, WB, dot-blot and IHC analyses were used to identify, quantify and validate dysregulation of proteins.
Results: Low C1GALT1 protein expression levels associate to a more aggressive phenotype of EC. Out of 5208 proteins identified and quantified by LC-MS/MS, 100 proteins showed dysregulation (log 2 fold-change ≥ 0.58 or ≤-0.58) in the cell protein extracts and 144 in the secretome of C1GALT1 depleted ECC-1 cells. Nine dysregulated proteins were validated. Bioinformatics analyses pointed out to an increase in pathways associated with an aggressive phenotype. This finding was corroborated by loss-of-function cell-based assays demonstrating higher proliferation, invasion, migration, colony formation and angiogenesis capacity in C1GALT1 depleted cells. These effects were associated to the overexpression of ANXA1, as demonstrated by ANXA1 transient silencing cell-based assays, and thus, correlating C1GALT and ANXA1 protein expression and biological effects. Finally, the negative protein expression correlation found by proteomics between C1GALT1 and LGALS3 was confirmed by IHC.
Conclusion: C1GALT1 stably depleted ECC-1 cells mimic an EC aggressive phenotype observed in patients and might be useful for the identification and validation of EC markers of progression.
(© 2023. The Author(s).)
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معلومات مُعتمدة: FPU predoctoral contract to A.M-C. Ministerio de Educación, Cultura y Deporte; PI17/01723 AES-ISCIII; PI20CIII/00019 AES-ISCIII
فهرسة مساهمة: Keywords: C1GALT1; Endometrial cancer; O-glycosylation; Quantitative proteomics; SILAC
المشرفين على المادة: EC 2.4.1.- (C1GALT1 protein, human)
EC 2.4.1.- (Galactosyltransferases)
تواريخ الأحداث: Date Created: 20230206 Date Completed: 20230525 Latest Revision: 20230526
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC10205863
DOI: 10.1007/s13402-023-00778-w
PMID: 36745330
قاعدة البيانات: MEDLINE
الوصف
تدمد:2211-3436
DOI:10.1007/s13402-023-00778-w