Extracellular Vesicles Derived From Lipopolysaccharide-Challenged Gingival Fibroblast Reveal Distinct miRNA Expression Patterns Associated With Reduced Cancer Survival

Abstract

Objectives

Periodontitis is a prevalent inflammatory disease with established systemic implications. Extracellular vesicles (EVs) have emerged as key mediators of intercellular communication, potentially linking periodontitis to systemic diseases. However, the molecular cargo of EVs from inflamed periodontal cells remains poorly characterized. This study investigates the EV cargo of human gingival fibroblasts (hGF-hTERT) following lipopolysaccharide (LPS) stimulation and explores their potential role in cancer progression.

Materials and Methods

EVs were isolated from LPS-treated and untreated fibroblasts via ultracentrifugation. Dynamic light scattering and scanning electron microscopy characterized EV size and morphology. RNA sequencing identified differentially expressed miRNAs, validated by qPCR. Functional pathway enrichment and in-silico analyses using The Cancer Genome Atlas (TCGA) were performed to assess EV-associated miRNA impact on tumorigenesis.

Results

EV size and concentration remained unchanged after LPS stimulation. However, LPS-derived EVs exhibited a 2.6-fold increase in miRNA content, with three significantly upregulated miRNAs: miR-146a-5p, miR-486-5p, and miR-451a. Functional enrichment analysis revealed their involvement in inflammation, immune modulation, and cancer pathways. In vitro, LPS-derived EVs significantly enhanced prostate cancer (LnCap) cell proliferation. TCGA analysis linked the upregulated miRNAs to poor cancer prognosis.

Conclusions

LPS stimulation alters the miRNA cargo of gingival fibroblast-derived EVs, enhancing pathways associated with inflammation and cancer progression. These findings suggest a mechanistic role for periodontal EVs in systemic disease pathogenesis, warranting further investigation into their diagnostic and therapeutic potential.