Effect Of ?-particle Radiation On Micro Rna Expression Profile Of Human Bronchial Epithelial Cells

Objective: Human bronchial epithelial cells(BEAS-2B)were used as the research object to investigate the effect of cumulative irradiation of ?-particles on the micro RNA expression profile of bronchial epithelial cells,and to screen potential markers of bronchial epithelial cell damage induced by ?-particle radiation,revealing possible regulatory mechanisms of bronchial epithelial cells of ?-particle radiation-induced.Merhods:(1)The normal bronchial epithelial cells BEAS-2B were irradiated by the 241 Am ? source of Soochow University.The subculture was performed by sub-cumulative irradiation.The doses were 0.1Gy,0.5Gy,and 2.0Gy,and the unirradiated cells were used as the control group.After each irradiation,the cells were subcultured for 10 passages,accumulated for 4 times,and subcultured for 40 passages.The established cell model was labeled 2B-n Gym(n represents the dose per exposure and m represents the number of e×posures).The final cell model was obtained as follows:2B-0.1Gy-4,2B-0.5Gy-4,2B-2.0Gy-4 and 2B-NC-4.Through cell migration and invasion ability detection,cell proliferation ability analysis,it is evaluated whether the ?-particle radiation-induced BEAS-2B cell damage model is successfullyconstructed.(2)Collected cumulative irradiation 4 times,subcultured 40 passages of cell models 2B-0.1Gy-4,2B-0.5Gy-4,2B-2.0Gy-4and2B-NC-4,micro RNA microarray was used for micro RNA expression profiling to screening differentially expressed micro RNA.(3)The target gene is predicted by differentially expressed micro RNA,and further analyzed by bioinformatics such as GO function annotation and KEGG pathway,to revealing the potential regu Lation mechanism for damage of ?-particle radiation-induced of bronchial epithelial cell.Results:(1)The ability of cell migration and invasion,cell proliferation were analyzed by cumulative irradiation 4 times and subcultured for 40 passages of cell model.Finally,the BEAS-2B cell injury model induced by ?-particle irradiation was obtained.Transwell experiments show that:compared with the control group,the migration and invasion ability of each treatment group is enhanced and related to dose.The cell proliferation ability of CCK-8 showed:that compared with the control,the proliferation ability of the cells in each experimental group increased significantly.(2)Micro RNA microarray results showed that:long-term accumulation of ?-particle radiation can induced differential expression of micro RNA in BEAS-2B cells.Among them,the 2B-0.1Gy-4 experimental group obtained 81 differentially expressed micro RNA,43 were up-regulated and 38 were downregulated;2B-0.5Gy-4 experimental group obtained 196 differentially expressed micro RNA,86 were up-regulated and 110 were down-regulated;In the 2B-2.0Gy experimental group,141 differentially expressed micro RNA were obtained,50 were up-regulated and 91 were down-regulated.In the three dose groups,42 micro RNA were differentially expressed,14 were up-regulated and 18 were down-regulated.Three dose groups were randomly selected to express the significant micro RNA mi R-4257,mi R-224,mi R-652-3p and mi R-4646-5p for real-time quantitative PCR analysis.The resu Lts were consistent with the chip.(3)Target Scan,PITA,and micro RNAorg were used to predict target genes of differential micro RNA.The GO function annotation and KEGG pathway bioinformatics analysis showed that the 2B-0.1Gy-4 differential gene mainly involved cell growth and development,ion transport,cell adhesion,cell inflammatory and other processes;2B-0.5Gy-4 differential gene mainly involves transmembrane transport,ion transport,nervous system growth anddevelopment,positive correlation regllation of cell proliferation;2B-2.0Gy-4 differential gene mainly involves DNA replication regulation,signal transduction,perception of apoptosis,cell cycle arrest,and response to DNA damage;With the increase of dose,2B-0.5Gy-4 and 2B-2.0Gy-4,showed PI3-Akt signaling pathway and ras signaling pathway related to cell malignant transformation increased,suggesting that the degree of cell damage is aggravated with the addition of dose.Conclusions:(1)The damaged cell model of human bronchial epithelial cells BEAS-2B induced by ?-particle irradiation was successfully constructed by means of fractional cumulative irradiation.(2)?-particle irradiation induces differential expression of micro RNA in BEAS-2B cells.Among them,mi R-4257,mi R-224,mi R-652-3p,and mi R-4646-5p showed consistent trend of differential expression in each dose group,and it is expected to be a molecular marker for radiation damage of bronchial epithelial cells induced by ?-particle.(3)Bioinformatics analysis of differentially expressed micro RNA showed that low dose radiation irradiation induced cell interaction and inflammatory response-related pathways,and as the irradiation dose increased,the enrichment of cancer and other related pathways appeared,and the number of pathways related to transformation adds,revealing that with the irradiation dose degree of cell damage increased.