Cancer-Associated Fibroblasts Promote Migration and Invasion of Non-Small Cell Lung Cancer Cells via miR-101-3p Mediated VEGFA Secretion and AKT/eNOS Pathway

Abstract

Cancer-associated fibroblasts (CAFs) are major component of tumor microenvironment (TME), which plays crucial roles in tumor growth, invasion and metastasis; however, the underling mechanism is not fully elucidated. Despite many studies are focused on the tumor promoting effect of CAFs-derived cytokines, the upstream regulators of cytokine release in CAFs is largely unknown. Here we found that miR-101-3p was downregulated in primary lung cancer-associated CAFs compared to normal fibroblasts (NFs). Ectopic overexpression of miR-101-3p suppressed CAFs activation, and abrogated the promoting effect of CAFs on migration and invasion of non-small cell lung cancer cells (NSCLC), through attenuating CAFs’ effect on epithelial mesenchymal transition (EMT) process, metastasis-related genes (MMP9, TWIST1) and AKT/endothelial nitric oxide synthase (eNOS) signaling pathway. Further study indicated that vascular endothelial growth factor A (VEGFA) was a novel target of miR-101-3p, and CAFs-derived VEGFA mediated the effect of miR-101-3p on migration and invasion of lung cancer cells, demonstrated by using recombinant VEGFA and VEGFA neutralizing antibody. Interestingly, the analysis of the Cancer Genome Atlas (TCGA) database revealed that lung cancer tissues expressed lower level of miR-101-3p than non-cancerous tissues, and low/medium-expression of miR-101-3p was associated with poor overall survival (OS) rate. Moreover, the mouse xenograft experiment also showed that CAFs accelerated tumor growth whereas miR-101-3p diminished CAFs’ effect. These findings revealed a novel mechanism that CAFs facilitated lung cancer metastasis potential via miR-101-3p/VEGFA/AKT signaling pathway, suggesting miR-101-3p as a potential candidate for metastasis therapy.

AZD6738 Inhibits fibrotic response of conjunctival fibroblasts by regulating checkpoint kinase 1/P53 and PI3K/AKT pathways

“…Therefore, the down-regulation of MMP9 in vitro under the action of different doses of AZD6738 verified the dual pharmacological mechanism of AZD6738 against fibrosis, which was caused by the negative feedback caused by the down-regulation of the target proteins like COL4 and COL5 ( Tschesche et al, 1992 ). Further, we revealed that AZD6738 reduces the expression of VEGFA in HConFs possible by down-regulating the phosphorylation level of eNOS through AKT / eNOS pathway ( Guo et al, 2021 ; Ninchoji et al, 2021 ), which shows the potential of AZD6738 in the treatment of neovascular eye diseases. Although TGF-β1 did not induce the up-regulation of RhoA level in HConFs in our research, the administration of high-dose AZD6738 still reduced the expression of RhoA to a certain extent, which indicating the potential effect of AZD6738 on the cytoskeleton.…”

Cancer-associated fibroblasts promote migration and invasion of non-small cell lung cancer cells via METTL3-mediated RAC3 m6A modification

“…The ability of cell migration was assessed by the wound healing assay as previously described 18 . Cells were plated on 6-well plates, pretreated with 0.05 μM mitomycin C, and cultured under different conditions.…”

MiR-101: An Important Regulator of Gene Expression and Tumor Ecosystem

“…Ectopic overexpression of miR-101 suppressed cancer-associated fibroblasts (CAFs) activation and abrogated the promoting effect of CAFs on migration and invasion of non-small cell lung cancer cells (NSCLC) through attenuating CAFs’ effect on epithelial–mesenchymal transition (EMT) process, metastasis-related genes matrix metallopeptidase 9 (MMP9), twist family BHLH transcription factor 1(TWIST1-), and AKT/endothelial nitric oxide synthase (eNOS) signaling pathway. Further study indicates that vascular endothelial growth factor A (VEGFA) is a novel target of miR-101, and CAFs-derived VEGFA mediates the effect of miR-101 on migration and invasion of lung cancer cells [ 61 ], suggesting a new combination of VEGF and miR-101 for metastasis therapy. Ceramide synthesis by Ceramide Synthase 6 (CERS6) is required for cell migration and metastasis in lung cancer.…”