Abstract
Numerous epidemiological studies have demonstrated that chronic PM2.5 exposure was associated with the lung carcinogenesis without known potential mechanisms. Exosomes-derived non-coding RNAs, including miRNAs, are proposed to play critical role in the occurrence and development of malignant diseases. So identification of exosomes-derived miRNAs could help us to better understand the molecular toxicity of PM2.5-induced lung cancer. Establishment chronic exposure animal and cell model with PM2.5 was conducted as before. HE staining was used for estimating the histological alternations of lungs in vivo. The expressions of EMT markers in vivo and vitro were quantified by Western blot. Then the exosomes in cell culture supernatant were extracted and the involved miRNAs were extracted and sequenced. The different expression level of miRNAs were verified by RT-PCR. Chronic PM2.5 exposure induced bronchial epithelial cell atypical hyperplasia and massive macrophage infiltration. PM2.5 exposure induce EMT event in vivo and vitro indicated as increased expression of Vimentin and decreased expression of E-cadherin. And five passages of PM2.5 stimulation also induced the release of rich and extractable exosomes in the cell culture supernatant in vitro. Through sequencing, there were differentially expressed 36 miRNAs between PM2.5 chronic exposed and control groups with 1.5-fold and greater differences. Among them, there were 30 exosome-miRNAs upregulated and 6 downregulated expression by PM2.5 exposure. The downregulated expression of miR-29b-2-5p, miR-193b-5p and miR-320c and upregulated expression of miR-100-5p, 125b-5p and unconservative_2_45093 in PM2.5 group were identified and reconfirmed by qRT-PCR. Chronic PM2.5 exposure causes bronchial epithelial cells atypical hyperplasia and induces EMT event in vivo, and it also induce the expression differences of miRNAs in exosome in vitro. Meanwhile, the identified differentially expressed exosome-miRNAs may partially associate with tumorigenesis. To sum up, the identified exosome-miRNAs may play role in the development of lung cancer induced by chronic PM2.5 exposure.