Carcinoembryonic antigen is a sialyl Lewis x/a carrier and an E‑selectin ligand in non‑small cell lung cancer

Ferreira,Inês   Gomes; Carrascal,Mylène; Mineiro,A.   Gonçalo; Bugalho,António; Borralho,Paula; Silva,Zélia; Dall'olio,Fabio; Videira,Paula  A.

doi:10.3892/ijo.2019.4886

Carcinoembryonic antigen is a sialyl Lewis x/a carrier and an E‑selectin ligand in non‑small cell lung cancer

Authors:
- Inês Gomes Ferreira
- Mylène Carrascal
- A. Gonçalo Mineiro
- António Bugalho
- Paula Borralho
- Zélia Silva
- Fabio Dall'olio
- Paula A. Videira
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Affiliations: Department of Experimental Diagnostic and Specialty Medicine, University of Bologna, Bologna I‑40126, Italy, CEDOC, NOVA Medical School, NOVA University of Lisbon, Lisbon 1150‑082, Portugal, UCIBIO, Department of Life Sciences, Faculty of Sciences and Technology, NOVA University of Lisbon, Caparica 2829‑516, Portugal, Department of Anatomical Pathology, Faculty of Medicine, University of Lisbon, Lisbon 1649‑028, Portugal
Published online on: September 26, 2019 https://doi.org/10.3892/ijo.2019.4886
Pages: 1033-1048
Copyright: © Ferreira et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The formation of distant metastasis resulting from vascular dissemination is one of the leading causes of mortality in non‑small cell lung cancer (NSCLC). This metastatic dissemination initiates with the adhesion of circulating cancer cells to the endothelium. The minimal requirement for the binding of leukocytes to endothelial E‑selectins and subsequent transmigration is the epitope of the fucosylated glycan, sialyl Lewis x (sLex), attached to specific cell surface glycoproteins. sLex and its isomer sialyl Lewis a (sLea) have been described in NSCLC, but their functional role in cancer cell adhesion to endothelium is still poorly understood. In this study, it was hypothesised that, similarly to leukocytes, sLe glycans play a role in NSCLC cell adhesion to E‑selectins. To assess this, paired tumour and normal lung tissue samples from 18 NSCLC patients were analyzed. Immunoblotting and immunohistochemistry assays demonstrated that tumour tissues exhibited significantly stronger reactivity with anti‑sLex/sLea antibody and E‑selectin chimera than normal tissues (2.2‑ and 1.8‑fold higher, respectively), as well as a higher immunoreactive score. High sLex/sLea expression was associated with bone metastasis. The overall α1,3‑fucosyltransferase (FUT) activity was increased in tumour tissues, along with the mRNA levels of FUT3, FUT6 and FUT7, whereas FUT4 mRNA expression was decreased. The expression of E‑selectin ligands exhibited a weak but significant correlation with the FUT3/FUT4 and FUT7/FUT4 ratios. Additionally, carcinoembryonic antigen (CEA) was identified in only 8 of the 18 tumour tissues; CEA‑positive tissues exhibited significantly increased sLex/sLea expression. Tumour tissue areas expressing CEA also expressed sLex/sLea and showed reactivity to E‑selectin. Blot rolling assays further demonstrated that CEA immunoprecipitates exhibited sustained adhesive interactions with E‑selectin‑expressing cells, suggesting CEA acts as a functional protein scaffold for E‑selectin ligands in NSCLC. In conclusion, this work provides the first demonstration that sLex/sLea are increased in primary NSCLC due to increased α1,3‑FUT activity. sLex/sLea is carried by CEA and confers the ability for NSCLC cells to bind E‑selectins, and is potentially associated with bone metastasis. This study contributes to identifying potential future diagnostic/prognostic biomarkers and therapeutic targets for lung cancer.

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