High levels of Notch intracellular cleaved domain are associated with stemness and reduced bevacizumab efficacy in patients with advanced colon cancer

Negri,Francesca; Bozzetti,Cecilia; Pedrazzi,Giuseppe; Azzoni,Cinzia; Bottarelli,Lorena; Squadrilli,Anna; Lagrasta,Costanza; Tamagnini,Ione; Bisagni,Alessandra; Ragazzi,Moira; Porzio,Rosa; Tomasello,Gianluca; Mori,Daniele; Leonardi,Francesco; Gnetti,Letizia; Crafa,Pellegrino; Sala,Roberto; Cascinu,Stefano

doi:10.3892/or.2019.7349

High levels of Notch intracellular cleaved domain are associated with stemness and reduced bevacizumab efficacy in patients with advanced colon cancer

Authors:
- Francesca Negri
- Cecilia Bozzetti
- Giuseppe Pedrazzi
- Cinzia Azzoni
- Lorena Bottarelli
- Anna Squadrilli
- Costanza Lagrasta
- Ione Tamagnini
- Alessandra Bisagni
- Moira Ragazzi
- Rosa Porzio
- Gianluca Tomasello
- Daniele Mori
- Francesco Leonardi
- Letizia Gnetti
- Pellegrino Crafa
- Roberto Sala
- Stefano Cascinu
View Affiliations

Affiliations: Medical Oncology Unit, University Hospital of Parma, I‑43126 Parma, Italy, Department of Medicine and Surgery, Unit of Neuroscience and Robust Statistics Academy (Ro.S.A.), University of Parma, I‑43126 Parma, Italy, Department of Medicine and Surgery, Unit of Pathological Anatomy, University Hospital of Parma, I‑43126 Parma, Italy, Pathology Unit, Istituto di Ricerca e Cura a Carattere Scientifico (IRCCS), Santa Maria Nuova Hospital, I‑42123 Reggio Emilia, Italy, Department of Oncology‑Haematology, G. da Saliceto Hospital, I‑29121 Piacenza, Italy, Medical Oncology Unit, ASST Ospedale di Cremona, I‑26100 Cremona, Italy, Department of Medicine and Surgery, General Pathology, University of Parma, I‑43126 Parma, Italy, Division of Medical Oncology, University Hospital of Modena and Reggio Emilia, I‑41124 Modena, Italy
Published online on: October 1, 2019 https://doi.org/10.3892/or.2019.7349
Pages: 2750-2758

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Abstract

δ‑like ligand 4 (DLL4)‑Notch signaling is associated with tumor resistance to anti‑vascular endothelial growth factor (VEGF) therapy. Furthermore, Notch signaling is critical for the maintenance of colon cancer stem cells (CSCs), which are relevant in drug resistance and tumor angiogenesis. CD44 is a transmembrane glycoprotein and is considered a putative marker of CSCs. To assess the association of Notch intracellular cleaved domain (NICD), DLL4 and CD44 expression with the efficacy of anti‑angiogenic drugs, a series of samples derived from patients with advanced colon cancer enrolled in prospective clinical trials were analyzed. Histological samples from 51 primary tumors that originated from patients treated with bevacizumab‑based first‑line chemotherapy were analyzed by immunohistochemistry for NICD, DLL4 and CD44 expression, and CD31 for microvessel count. The expression levels of genes relevant for angiogenesis [angiopoietin (ANGPT)1, ANGPT2, fibroblast growth factor (FGF)1, FGF2, epidermal growth factor, placental growth factor, VEGFA and DLL4] were detected by reverse transcription‑quantitative PCR using RNA extracted from the frozen tissues of four tumors with low and four tumors with high NICD expression. Strong NICD levels were observed in 12/51 (24%) of the patients, whereas 16/51 (31%) of the colon cancer subjects exhibited high CD44 expression. Strong CD44 staining was associated with high NICD levels compared with the CD44 expression levels noted in samples with low NICD levels (67 vs. 20%, P=0.005). No association was observed with regards to the expression levels of NICD, CD44 and the other aforementioned biomarkers. High expression levels of NICD and CD44 predicted reduced progression‑free survival (P<0.001) and overall survival (P=0.002). No significant differences in the expression of angiogenesis‑related genes were detected between low and high NICD‑expressing tumors. In conclusion, NICD and CD44 tissue levels exhibited an association and may be related to a reduced survival rate in patients with advanced colon cancer treated with bevacizumab.

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