Mechanism Of EMT Changes In Bronchial Epithelial Cells Induced By Ionizing Radiation

Objective:In order to explore the mechanism of this gene in human bronchial epithelial mesenchymal transition(EMT)induced by long-term low-dose ionizing radiation,differentially expressed genes in the transcriptome of radiation workers are screened and searched for key genes.Methods:1.RNA-seq transcriptomics are used to detect blood samples from a unit of radiation workers,bioinformatics analysis of RNA-seq results,screening for differentially expressed mRNA and verification by real-time fluorescent PCR(RT-PCR).2.X-ray irradiator was used to irradiate human bronchial epithelial cells(16HBE)multiple times,with a single dose of 0.5Gy and a cumulative dose of 2Gy.3.The irradiated cells were tested for immunofluorescence,scratch healing,cell adhesion,and CCK-8 activity to verify the presence of epithelial-mesenchymal transition(EMT).4.Western Blot experiments were used to verify the expression of FN1 protein in cells with different cumulative doses,and to determine whether FN 1 could be used as a biomarker for the induction of EMT in normal bronchial epithelial cells induced by ionizing radiation.Results:1.Transcriptome results showed that a total of 873 mRNAs had significant differences,of which 68 were up-regulated and 805 were down-regulated.Differentially expressed genes are mostly related to biological processes such as epithelial-mesenchymal transition(EMT),immune response,and cell migration.2.With the increase of cumulative dose of human normal bronchial epithelial cells(16HBE),the expression of cadherin(E-cad)gradually decreased and vimentin(Vimentin)gradually increased.The cell migration ability is enhanced,the adhesion rate is reduced,and preliminary EMT is formed.3.The results of WesternBlot experiments found that as the cumulative dose increased,the expression of FN1 in cells gradually increasedConclusion:This subject performs high-throughput sequencing and analysis of blood samples from radiation workers,and screens the differential mRNA expression profiles of radiation workers and exposed workers.Bronchial epithelial cells were irradiated and ionizing radiation was established to induce EMT.It was found that FN1 protein can be used as a new biomarker of radiation-induced EMT.