Profiling and Functional Analysis of microRNA Deregulation in Cancer-Associated Fibroblasts in Oral Squamous Cell Carcinoma Depicts an Anti-Invasive Role of microRNA-204 via Regulation of Their Motility

Abstract

Background: Knowledge on the role of miR changes in tumor stroma for cancer progression is limited. This study aimed to investigate the role of miR dysregulation in cancer-associated fibroblasts (CAFs) in oral squamous cell carcinoma (OSCC). Methodology: CAF and normal oral fibroblasts (NOFs) were isolated from biopsies of OSCC patients and healthy individuals after informed consent and grown in 3D collagen gels. Total RNA was extracted. Global miR expression was profiled using Illumina version 2 panels. The functional impact of altered miR-204 expression in fibroblasts on their phenotype and molecular profile was investigated using mimics and inhibitors of miR-204. Further, the impact of miR-204 expression in fibroblasts on invasion of adjacent OSCC cells was assessed in 3D-organotypic co-cultures. Results: Unsupervised hierarchical clustering for global miR expression resulted in separate clusters for CAF and NOF. SAM analysis identified differential expression of twelve miRs between CAF and NOF. Modulation of miR-204 expression did not affect fibroblast cell proliferation, but resulted in changes in the motility phenotype, expression of various motility-related molecules, and invasion of the adjacent OSCC cells. 3′ UTR miR target reporter assay showed ITGA11 to be a direct target of miR-204. Conclusions: This study identifies differentially expressed miRs in stromal fibroblasts of OSCC lesions compared with normal oral mucosa and it reveals that one of the significantly downregulated miRs in CAF, miR-204, has a tumor-suppressive function through inhibition of fibroblast migration by modulating the expression of several different molecules in addition to directly targeting ITGA11.

Red blood cell-derived extracellular vesicles with miR-204 mimic loading for pediatric neuroblastoma treatment

“…RBC-EV miR-204 decreased cell viability, mitigated cell migration, and suppressed spheroid formation and growth, more strongly against MYCN-amp NB cells, but these anti-neuroblastoma effects were also observed, but to a lesser extent, in MYCN-NA cells (Figures 5-7). These results are consistent with previous evidence, in which miR-204 exhibited anticancer activities against several cancers with diversed oncogenic backgrounds, i.e., oral squamous cell carcinoma (OSCC), renal cell carcinoma, and non-small-cell lung cancer [47][48][49][50]. As RBC-EV miR-204 could inhibit both NB cell migration and spheroid formation and growth, the use of RBC-EV miR-204 as an adjuvant therapy may mitigate metastasis in high-risk neuroblastoma, regardless MYCN amplification status.…”

Effects of cancer‐associated fibroblasts deletion using HSVtk suicide system in OSCC

“…Cancer-associated fibroblasts (CAFs) are one major sub-cell type within the tumor microenvironment that has drawn attention. Multiple studies have confirmed that the increasing presence of CAFs associates poorly with prognosis in lung, prostate, and oral cancer (Knops et al, 2020;Rajthala et al, 2021;Wu et al, 2021). Specifically, CAFs are reported to promote cell proliferation, regulate immune responses, remodel ECM, and induce drug resistance by cytokine and metabolite secretion as well as extracellular vesicle transfer (Gieniec et al, 2019;Levi-Galibov et al, 2020).…”

Fibroblasts as Turned Agents in Cancer Progression

“…mir-34a-5p expression is significantly decreased in CAFs, resulting in a loss of its tumor suppressive effect [ 120 ]. The expression of miR-204 in cancer-adjacent fibroblasts also inhibits tumors by suppressing fibroblast migration by modulating the expression of signaling molecules and by directly targeting integrin α11 (ITGA11) and is also downregulated in HNSSC of the oral cavity [ 121 ].…”