The critical effects of matrices on cultured carcinoma cells: Human tumor-derived matrix promotes cell invasive properties

Abstract

The interaction between squamous cell carcinoma (SCC) cells and the tumor microenvironment (TME) plays a major role in cancer progression. Therefore, understanding the TME is essential for the development of cancer therapies.We used four (primary and metastatic) head and neck (HN) SCC cell lines and cultured them on top of or within 5 matrices (mouse sarcoma-derived Matrigel®, rat collagen, human leiomyoma-derived Myogel, human fibronectin, and human fibrin). We performed several assays to study the effects of these matrices on the HNSCC behavior, such as proliferation, migration, and invasion, as well as cell morphology, and molecular gene profile.Carcinoma cells exhibited different growth patterns depending on the matrix. While fibrin enhanced the proliferation of all the cell lines, collagen did not. The effects of the matrices on cancer cell migration were cell line dependent. Carcinoma cells in Myogel-collagen invaded faster in scratch wound invasion assay. On the other hand, in the spheroid invasion assay, three out of four cell lines invaded faster in Myogel-fibrin. These matrices significantly affected hundreds of genes and a number of pathways, but the effects were cell line dependent.The matrix type played a major role in HNSCC cell phenotype. The effects of the ECMs were either constant, or cell line dependent. Based on these results, we suggest to select the most suitable matrix, which provides the closest condition to the in vivo TME, in order to get reliable results in in vitro experiments.

Transcriptomic Profiling of JEG-3 cells using human leiomyoma derived matrix

“…Adding PDMS on Myogel + fibrin coating showed no obvious changes in cell morphology and no organoid structures, but 36 genes were up-regulated. Collectively, these morphological findings are in line with the previous observations, showing that Myogel provides better cell migration, and Myogel + fibrin coating generates two and three dimensional matrices [24] , [25] , [36] . Considered together, these data indicate that the coating has a marked impact on gene expression and morphology, including key cellular processes such as ribosome biosynthesis (mainly Myogel related effect), endocytosis, and adhesion-associated genes such as several of the matrix metalloproteases [37] , [38] .…”

“…Normalization and differential expression analysis between these two data sets were performed using edgeR [32] package integrated into Chipster platform [33] . Heatmaps were drawn with heatmapper tool [34] and pathway analyses were performed with Enrichr (MSigDB Hallmark 2020 pathway) [35] , [36] .…”

A 3D culture system for evaluating the combined effects of cisplatin and anti-fibrotic drugs on the growth and invasion of lung cancer cells co-cultured with fibroblasts

“… 28 Moreover, the gene profiles of cells cultured in fibrin or fibrin/Matrigel might be different from those of cells in Matrigel. 29 Because pure fibrin might not resemble the ECM constituents in the TME, we decided to use fibrin/Matrigel in the following experiments.…”

Modeling the Role of Cancer-Associated Fibroblasts in Tumor Cell Invasion

“…Conveniently, the aforementioned assays can be modified to incorporate a matrix into their set-up, allowing researchers to easily study both biological processes without investing in extra equipment ( Figure 1 A, top and middle row). In the transwell system, the porous membrane is coated with a layer of the matrix before seeding cells on top [ 21 ], whereas in both the scratch wound and exclusion zone systems, a layer of the matrix is added to embed the cells after the cell-free area has been established in the monolayer [ 22 ]. In both cases, collagen or preparation of a basement membrane matrix such as Matrigel (Corning) or Cultrex (R&D) are the most commonly used.…”