Cancer-Associated Fibroblasts Modulate Transcriptional Signatures Involved in Proliferation, Differentiation and Metastasis in Head and Neck Squamous Cell Carcinoma

Abstract

Cancer-associated fibroblasts (CAFs) are known to increase tumor growth and to stimulate invasion and metastasis. Increasing evidence suggests that CAFs mediate response to various treatments. HNSCC cell lines were co-cultured with their patient-matched CAFs in 2D and 3D in vitro models, and the tumor cell gene expression profiles were investigated by cDNA microarray and qRT-PCR. The mRNA expression of eight candidate genes was examined in tumor biopsies from 32 HNSCC patients and in five biopsies from normal oral tissue. Differences in overall survival (OS) were tested with Kaplan–Meier long-rank analysis. Thirteen protein coding genes were found to be differentially expressed in tumor cells co-cultured with CAFs in 2D and 81 in 3D when compared to tumor cells cultured without CAFs. Six of these genes were upregulated both in 2D and 3D (POSTN, GREM1, BGN, COL1A2, COL6A3, and COL1A1). Moreover, two genes upregulated in 3D, MMP9 and FMOD, were significantly associated with the OS. In conclusion, we demonstrated in vitro that CAF-derived signals alter the tumor cell expression of multiple genes, several of which are associated with differentiation, epithelial-to-mesenchymal transition (EMT) phenotype, and metastasis. Moreover, six of the most highly upregulated genes were found to be overexpressed in tumor tissue compared to normal tissue.

Isolation and characterization of head and neck cancer-derived peritumoral and cancer-associated fibroblasts

“…Conditioned media of CAFs promoted proliferation of Cal27, Cal33, and FaDu carcinoma cells, which was significantly higher than effects of hOFs ( Figure 4 ). A proliferation-inducing capacity of CAFs is in line with the general view of growth-promoting effects of tumor-resident CAFs ( 13 , 49 , 50 ). Effects of CAFs and PtFs on cancer cell migration was assessed by in vitro measurements.…”

Disruption of pancreatic stellate cell myofibroblast phenotype promotes pancreatic tumor invasion

“…First, we profiled the expression status of 411 gene transcripts associated with the ECM and cell adhesion (QIAGEN). WT and PKN2 KO PSCs were cultured in complete medium or exposed to TGF-β1 for 72 h. PKN2 deletion predominantly resulted in the upregulation of ECM-associated genes under 2D cell culture conditions ( Figures 2 A and 2B), including genes associated with metastasis (Serpine2, Fmod, Itgbl1, Aspn, MMP28, and Col6a3; Buchholz et al., 2003 ; Liu et al., 2021 ; Wiechec et al., 2021 ). Many of these genes were also differentially expressed (DE) in PKN2 KO PSCs compared with WT PSCs following TGF-β1 stimulation ( Figures 2 B and 2C; Table S1 ).…”

NT5E upregulation in head and neck squamous cell carcinoma: A novel biomarker on cancer-associated fibroblasts for predicting immunosuppressive tumor microenvironment

“…During the progress of ECM remodeling, the deposit of collagens IV, VII, XI, and XV is promoted (17), and thus the ECM contractility and stromal stiffness are augmented by CAFs, which is correlated to promoted tumor malignancy and poor patient prognosis (18). Besides, CAFs can secrete transforming growth factor-b (TGF-b), epidermal growth factor receptor (EGFR), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), C-C chemokine ligand 2 (CCL2), C-C chemokine ligand 5 (CCL5), C-X-C chemokine ligand 12 (CXCL12), and other active factors (19), and then act on tumor cells via paracrine or juxtacrine to activate a variety of important intracellular signal transduction pathways including TGF-b/Smad (20), PI3K/Akt/ mTOR (21), Wnt/b-catenin ( 22), IL-6/STAT3 (23) and Notch (24), thereby triggering epithelial-to-mesenchymal transition (EMT) in tumor cells (25). The expression of E-cadherin, tight junction protein 1 (ZO-1), cytokeratin (CK) and other cell adhesion molecules in epithelial tissues is inhibited, whereas the expression of N-cadherin, vimentin (VIM), a-smooth muscle actin (a-SMA/ACTA2), fibroblast specific protein 1 (FSP1/ S100A4) and Osteopontin in mesenchymal cells is induced (26), promoting the loss of epithelial apical-basal polarity, and driving the acquisition of a mesenchymal and motile phenotype (27).…”