Development and cytotoxic response of two proliferative MDA-MB-231 and non-proliferative SUM1315 three-dimensional cell culture models of triple-negative basal-like breast cancer cell lines.

Clémence Dubois,Christelle Blavignac,Claire Szczepaniak,Pierre Daumar,Corinne Aubel,Robin Dufour,Mahchid Bamdad,Emmanuelle Mounetou,Frédérique Penault-Llorca

doi:10.18632/oncotarget.20517

Abstract

Triple-Negative Basal-Like tumors, representing 15 to 20% of breast cancers, are very aggressive and with poor prognosis. Targeted therapies have been developed extensively in preclinical and clinical studies to open the way for new treatment strategies. The present study has focused on developing 3D cell cultures from SUM1315 and MDA-MB-231, two triple-negative basal-like (TNBL) breast cancer cell lines, using the liquid overlay technique. Extracellular matrix concentration, cell density, proliferation, cell viability, topology and ultrastructure parameters were determined. The results showed that for both cell lines, the best conditioning regimen for compact and homogeneous spheroid formation was to use 1000 cells per well and 2% Geltrex®. This conditioning regimen highlighted two 3D cell models: non-proliferative SUM1315 spheroids and proliferative MDA-MB-231 spheroids. In both cell lines, the comparison of 2D vs 3D cell culture viability in the presence of increasing concentrations of chemotherapeutic agents i.e. cisplatin, docetaxel and epirubicin, showed that spheroids were clearly less sensitive than monolayer cell cultures. Moreover, a proliferative or non-proliferative 3D cell line property would enable determination of cytotoxic and/or cytostatic drug activity. 3D cell culture could be an excellent tool in addition to the arsenal of techniques currently used in preclinical studies.

Highlights

Triple-negative basal-like (TNBL) tumors comprise 70% of the basal-like tumor subtype and 15% of all breast cancers
The present study has focused on developing 3D cell cultures from SUM1315 and MDA-MB-231, two triple-negative basal-like (TNBL) breast cancer cell lines, using the liquid overlay technique
The results showed that when Geltrex® was added just after cell seeding, loose cell aggregates were detected in the presence of 0.25 and 0.5% of Geltrex® concentrations (Table 1, Figure 1A)

Summary

Introduction

Triple-negative basal-like (TNBL) tumors comprise 70% of the basal-like tumor subtype and 15% of all breast cancers. TNBL tumors mainly affect young women and are frequently associated with hereditary predispositions (BRCA1/2 germline mutations). This subtype has a very poor prognosis. Conventional chemotherapy is based on different protocols such as FEC (5-fluorouracil, epirubicin, cyclophosphamide), FAC (5-fluorouracil, adriamycin, cyclophosphamide) or platinum salts (cisplatin), generally associated with side effects [5]. Otherwise, these tumors are not sensitive to classical breast targeted therapies since they do not express the relevant receptors (i.e. ER, PR and HER2). Targeting Epidermal Growth Factor Receptor 1 pathway with anti-EGFR Monoclonal antibodies (MoAb) or Tyrosine Kinase Inhibitors (TKI); or inhibiting the enzyme Poly-ADP-Ribose-Polymerase (reparation of single-strand breaks)(PARPi) have shown promising activities in this preclinical and clinical setting) [5,6,7,8]

Methods

Results

Conclusion