Study On Mechanism Of Cancer-related Gene Expression And Signaling Pathway In HBE Cells Treated By PM2.5

To study the mechanism of oncogene expression and the MAPK / PKC signaling pathway in human bronchial epithelial cells(HBE)exposed to PM2.5.Methods: HBE cells were treated with PM2.5 at 10 ?g / mL and 50?g / mL,and a control group was set up.10 ?M / L Cr6 + was used as a positive control.HBE cells were exposed for 24 h.Fluorescent quantitative PCR and Western blotting were used.)Detection of oncogene(c-myc,c-fos,k-ras and p53)gene and protein expression levels.MAPK / PKC signaling pathway experiments were set up with blank control group,PM2.5 experimental group,inhibitor group(p38MAPK,ERK1 / 2,PKC? inhibitor),and PM2.5 + inhibitor group.HBE cells were pretreated with inhibitors for 30 min,then cultured in serum-free DMEM medium for 24 h as the inhibitor experimental group,and PM2.5 was used for 24 h as the PM2.5 experimental group.HBE cells were pretreated with inhibitors for 30 min,and then treated with 50 ?g /mL PM2.5 for 24 h as the PM2.5 + inhibitor experimental group.Quantitative quantitative PCR and western blot were used to detect changes in mRNA and protein expression levels of p38 MAPK,ERK1,ERK2,PKC?,c-myc,c-fos,k-ras,and p53 genes.For proteomics experiments,HBE cells were treated with 50?g / mL PM2.5 samples from Shenzhen and Taiyuan.A control group was set up and the HBE cells were exposed for 24 h.Q Exactive mass spectrometer was used to detect the overall protein expression.Perseus software was used to screendifferential proteins.GO,KEGG analysis and Weighted Correlation Network analysis(WGCNA)were performed on the differential proteins.Results:1.Compared with the control group,the expression levels of c-myc gene in HBE cells of 10 ?g/mL and 50 ?g / ml PM2.5 group and the positive control group increased by 500.1%,780.7%,and 305.3%,respectively,and the expression level of c-fos gene increased by 34.0%,76.7%,131.3%,k-ras gene expression levels increased by 50.3%,107.0%,49.7%,p53 gene expression levels decreased by 28.3%,28.7%,and59.7%,the differences were statistically significant(P <0.05 or P<0.01).Compared with the control group,the expression levels of c-myc protein in HBE cells in the 50 ?g / ml PM2.5 group and the positive control group were increased by 29.7% and 77.3%,respectively;the expression levels of c-fos protein were increased by 200.3% and137.0%,respectively.All were statistically significant(P <0.05 or P<0.01).Compared with the control group,the expression levels of k-ras protein in HBE cells of 10 ?g/mL and 50 ?g / ml PM2.5 group and the positive control group increased by 106.3% and 130.3%,respectively.,116.7%,p53 protein expression levels decreased by 43.7%,53.3%,and52.1%,respectively,and the differences were statistically significant(P<0.05 or P <0.01).2.After treatment with p38 MAPK inhibitor SB203580,compared with the control group,the expression levels of p38 MAPK,c-myc,c-fos,and k-ras genes in the SB203580 experimental group decreased.Compared with the PM2.5 experimenta l group,in the PM2.5 +SB203580 experiment group,the expressions of p38 MAPK,c-myc,c-fos,and k-ras genes were all decreased,and the expression of p53 genes was increased in the group.After treatment with ERK1 / 2 inhibitor PD98059,compared with the control group,the expression of ERK1,ERK2,c-myc,c-fos,and k-ras in the PD98059 experimental group decreased.Compared with the PM2.5 experimental group,In the PM2.5 + PD98059 experimental group,the expressions of ERK1,ERK2,c-myc,c-fos,and k-ras all decreased,and the expression of p53 gene increased.After treatment with the PKC? inhibitor CHE,compared with the control group,the PKC? gene expression decreased in the CHE experimental group,and the c-myc,c-fos,k-ras,and p53 genes did not change significantly;compared with the PM2.5 experimental group,In the PM2.5 + CHE experimental group,the expressions of PKC?,c-myc,c-fos,and k-ras genes decreased,and the expression of p53 gene increased.3.By comparing the protein expression levels of the Shenzhen sample group and the Taiyuan sample group with the control group,a total of 67 differential proteins were selected in the Shenzhen sample processing group,of which 46 were up-regulated and 21 were down-regulated;the Taiyuan sample-treated group was screened in total There were 252 differential proteins,of which 134 were up-regulated and118 were down-regulated.After KEGG analysis,it was found that the differentially expressed proteins were mainly enriched in ubiquitin-mediated proteolysis and HIF-1 signal pathways after Shenzhen PM2.5 samples were exposed to HBE cells.The GO analysis results showed that the differential proteins caused by the Shenzhen sample were mainly involved in the biological process of the absorption of various metal ions and the cell components such as myelin sheath and main axon;Differential proteins are mainly involved in protein aggregation regulating biological processes,molecular function of oxidase activity,and cellular components such as mitochondria and ribosome.In addition,three valuable differentially expressed proteins were screened,including ANXA2,DIABLO,and AIMP1.Conclusion:1.Atmospheric fine particulate matter PM2.5 has a significant promotion effect on oncogene expression in HBE cells,andsmog carcinogenesis may be closely related to PM2.5 oncogene expression.2.PM2.5 can promote the expression of oncogenes including c-myc,c-fos and k-ras in HBE cells.The application of SB203580,PD98059 and CHE inhibitors can significantly reduce the effect of PM2.5 on oncogene expression.It is suggested that MAPK and PKC signaling pathways play an important role in PM2.5 oncogene expression.3.Shenzhen and Taiyuan PM2.5 samples have different effects on HBE cells in biological processes,molecular functions,and cell composition,providing a scientific basis for further research on the molecular mechanism of PM2.5 carcinogenesis.