Growth of MCF-7 breast cancer cells and efficacy of anti-angiogenic agents in a hydroxyethyl chitosan/glycidyl methacrylate hydrogel

Abstract

BackgroundBreast cancer negatively affects women’s health worldwide. The tumour microenvironment plays a critical role in tumour initiation, proliferation, and metastasis. Cancer cells are traditionally grown in two-dimensional (2D) cultures as monolayers on a flat solid surface lacking cell–cell and cell–matrix interactions. These experimental conditions deviate from the clinical situation. Improved experimental systems that can mimic the in vivo situation are required to discover new therapies, particularly for anti-angiogenic agents that mainly target intercellular factors and play an essential role in treating some cancers.MethodsChitosan can be modified to construct three-dimensional (3D) tumour models. Here, we report an in vitro 3D tumour model using a hydroxyethyl chitosan/glycidyl methacrylate (HECS–GMA) hydrogel produced by a series of chitosan modifications. Parameters relating to cell morphology, viability, proliferation, and migration were analysed using breast cancer MCF-7 cells. In a xenograft model, secretion of angiogenesis-related growth factors and the anti-angiogenic efficacy of Endostar and Bevacizumab in cells grown in HECS–GMA hydrogels were assessed by immunohistochemistry.ResultsHydroxyethyl chitosan/glycidyl methacrylate hydrogels had a highly porous microstructure, mechanical properties, swelling ratio, and morphology consistent with a 3D tumour model. Compared with a 2D monolayer culture, breast cancer MCF-7 cells residing in the HECS–GMA hydrogels grew as tumour-like clusters in a 3D formation. In a xenograft model, MCF-7 cells cultured in the HECS–GMA hydrogels had increased secretion of angiogenesis-related growth factors. Recombinant human endostatin (Endostar), but not Bevacizumab (Avastin), was an effective anti-angiogenic agent in HECS–GMA hydrogels.ConclusionsThe HECS–GMA hydrogel provided a 3D tumour model that mimicked the in vivo cancer microenvironment and supported the growth of MCF7 cells better than traditional tissue culture plates. The HECS–GMA hydrogel may offer an improved platform to minimize the gap between traditional tissue culture plates and clinical applicability. In addition, the anti-angiogenic efficacy of drugs such as Endostar and Bevacizumab can be more comprehensively studied and assessed in HECS–GMA hydrogels.

Chemical activation of prolyl hydroxylase-2 by BBAP-1 down regulates hypoxia inducible factor-1α and fatty acid synthase for mammary gland chemoprevention

“…The cells were counted using a trypan blue exclusion method in hemocytometer. 29 Cell viability assay ER + MCF-7 cells (1 Â 10 5 ) were seeded in 96-well sterile plates and treated with different concentrations of BBAP-1 (1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM) for 24 h against control and standard drug (tamoxifen). Aer 24 h of incubation at 37 C in an atmosphere of 5% CO 2 and 95% humidity, the apical and basolateral media were pooled on a 24-well plate and the cell viability was estimated by MTT assay through measuring the emitted uorescence with a spectrouorometric multi plate reader (excitation-555 nm, emission-585 nm).…”

A three dimensional in vivo model of breast cancer using a thermosensitive chitosan‐based hydrogel and 4 T1 cell line in Balb/c

“…The results showed a malignant phenotype in tumors created by the 3D cultured cells. Migration, cell growth, and tumor volume were higher in the 3D group 37 . In the present study, the creation of 3D model was different from previous studies in that 1 × 10 6 4 T1 cells (mouse breast cancer), after being harvested from 2D culture medium, was loaded in the hydrogel, and was injected into the flank of mice and the same number of cells were injected into the mice without hydrogel.…”

“…Migration, cell growth, and tumor volume were higher in the 3D group. 37 In the present study, the creation of 3D model was different from previous studies in that 1 × field. 38,39 Ultrasound examination of various types of human breast tumors revealed that the malignant tumor tissue is hyperecogenic, darker in sonography, and have higher tissue density, while benign tissue is hyperecogenic and its connective tissue and fat content is higher.…”

Self‐Assembled Peptide‐Based Biocomposites for Near‐Infrared Light Triggered Drug Release to Tumor Cells

“…This is similar to the morphologies of healthy MCF‐7 cells seen in previous works. [ 59 ] Cells in the presence of neat FLS assemblies (without drug, Figure S6b, Supporting Information) showed aggregation and appearance of ribbon‐like membrane ruffles upon adhesion to the assemblies. In previous studies, it has been reported that MCF‐7 cells, when treated with certain small molecules or drugs such as tamoxifen showed E‐cadherin‐dependent ability to form aggregates which in turn reduced invasive behavior, as MCF‐7/6 displays inverse relation between aggregation and invasion.…”