Implication Of Different Anti-Angiogenic Treatments On In Vitro Vascular Tube Formation In Triple Negative Inflammatory Breast Cancer Cell Lines.

Abstract Introduction: Inflammatory breast cancer (IBC) is the most aggressive breast cancer in women and female dogs (IMC, Inflammatory mammary cancer) with a poor prognosis. Both IBC and IMC are highly angiogenic, lymphangiogenic and lymphangioinvasive. The aim of this study was to evaluate the effect of different anti-angiogenic treatments in the in vitro formation of vascular-like structures as well as cell secretion of different angiogenic and lymphangiogenic factors (VEGFA, VEGFC, VEGFD) and interleukin-8 (IL-8) in culture media. Material and Methods: For in vitro tube formation assay, culture human SUM149 cells and canine IPC-366 cells were divided into a control group, treated with DMSO, and 4 experimental groups treated with 1.5 μM of VEGF, SU5416 (anti-VEGFR, VEGF B20-4.1.1 (anti-VEGF) and celecoxib (NSAID), respectively, for 4 h at 37ºC in 5% CO2 conditions. Images were taken every 2 hours. Incomplete networks were excluded, and vessel-like tube structures were counted using the Image J Angiogenesis Analyzer software. Data were represented as average percentage relative to control ± SD. Results: VEGF and celecoxib treatment significantly augmented the formation of vascular-like tube structures respect to control group in both cell lines. However, VEGF B20-4.1.1 and SU5416 decreased the formation of vascular-like tube structures but not significantly. Conditionate media of IPC-366 and SUM149-treated cells with VEGF had significantly decreased VEGFA levels. In contrast, celecoxib treatment, reduced VEGFD levels and increased VEGFC in both cell lines. VEGF B20-4.1.1 addition increased VEGFA levels in culture media and SU5416 significantly diminished VEGFD concentrations. No significant differences in IL-8 levels were found for any treatment. Conclusions: Increased VEGFC levels after celecoxib treatment suggest a switch from a proangiogenic state toward a lymphangiogenic state. Neutralization of VEGF by VEGF B20-4.1.1 enhances the secretion of VEGFA and VEGFC by tumor cells inhibiting angiogenesis and lymphangenesis. Citation Format: Sara Caceres, Angela Alonso-Diez, Belen Crespo, Eduardo ESpinosa, Beatriz Madaruiaga, Gema Silvan, Maria J. Illera, Juan Carlos Illera. Implication of different anti-angiogenic treatments on in vitro vascular tube formation in triple negative inflammatory breast cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2646.