Effects Of Nano-silica And Benzo[a]pyrene On TNF?-TNFR Signaling Pathway In Lung Cancer Of Xuanwei Women

Objectives:1.To investigate the effect of nano-silica and benzo[a]pyrene on the migration ability of BEAS-2B cells.2.To investigate the effect of nano-silica and benzo[a]pyrene on the invasive ability of BEAS-2B cells.3.To investigate the difference effects of combination or alone of nano-silica and benzo[a]pyrene on the TNF?-TNFR signaling pathway in three cell lines BEAS-2B,A549 and YTMLC4.To detect and analyze the differences in gene levels of TNF?-TNFR signaling pathway in female patients and non-Xuanwei female patients and male patients.Methods:1.Nanosilica and benzo[a]pyrene were used alone or in combination to infect BEAS-2B cells,and scratching experiments were performed on the infected cells.2.Nanosilica and benzo[a]pyrene were used alone or in combination to infect BEAS-2B cells,and transwell experiments were performed on the cells after exposure.3.Nano-silica and benzo[a]pyrene were used alone or in combination to infect BEAS-2B,A549,and YTMLC,RNA extracted from infected cells,and reverse-transcribed into CDNA.Three cell lines at three time points,P100,Caspase3,Caspase8,and TNF? genes were subjected to QPCR experiments.4.Collect the intraoperative cancer tissue samples and normal tissue specimens that meet the inclusion criteria,and use RT-QPCR method to detect the expression changes of P100,Caspase3,Caspase8,and TNF? key protein counterpart genes.5.Statistical analysis of data results using SPSS 22.0.Results:1.Scratch test:BEAS-2B cells were scratched 24 hours after exposure.Observation of the combined exposure group after 48 hours increased cell migration ability,and there was a statistically significant difference compared with other groups(P<0.05).2.Transwell test,BEAS-2B cell strains were infected 24 hours into the chamber,48 hours after the observation of the combined exposure group compared with the other groups have increased the number of penetrating cells,and the difference was statistically significant(P<0.05).3.Cell RTQPCR experiment,BEAS-2B cell group:(1)The expression level of P100 and TNFa corresponding genes in BEAS-2B cells after 12 and 24 hours of exposure was increased in the combination group,and the difference was statistically significant(3.P<0.05).(2)The expression of cas8 in 24 hours after exposure to BEAS-2B cells was increased in the combination group,and the difference was statistically significant(P<0.05).(3)There was no significant difference in TNF?,P100,Caspase3,and Caspase8 in BEAS-2B cells 48 hours after exposure.A549 cell group:(1)The expression levels of Caspase8,Caspase3,and TNF genes in A549 cells were decreased in the experimental group 12 hours after exposure to A549 cells,and there was a statistically significant difference compared with the control group(P<0.05);(2)A549 cells The expression levels of Caspase8,Caspase3,and TNF genes were increased in the experimental group 24 hours after exposure to the virus,and there were significant differences between the two groups compared with the control group(P<0.05).The combined exposure group and the single exposure were observed.There was a difference in the expression of Caspase8 in the group and a more significant increase in the combined group.The difference was statistically significant(P<0.05).(3)The expression of Caspase3 in A549 cells after 48 hours of exposure was decreased in the experimental group compared with the control group(P<0.05).YTMLC cell group:(1)The expression levels of Caspase8,Caspase3,and TNF genes in YTMLC cells were decreased in the experimental group after 12 hours of exposure to YTMLC cells,and the difference was statistically significant(P<0.05),and the expression level of Caspase8 gene was significantly different from the control group(P<0.05).The decline was most significant in the combined group,and the difference was statistically significant(P<0.05).(2)The expression of Caspase3 and TNF genes in YTMLC cells after 24 hours of exposure was decreased in the experimental group,and there was a statistically significant difference compared with the control group(P<0.05).(3)The expression levels of Caspase8,Caspase3,and TNF genes in YTMLC cells after 48 hours of exposure were all decreased in the experimental group,and there was a statistically significant difference compared with the control group(P<0.05).4.Tissue sample experiments,(1)The relative expression levels of Caspase3 and TNF? genes in the women of Xuanwei and non-Xuanwei female tissues were decreased,and the difference was statistically significant(P<0.05).(2)There was no significant difference in the expression of P100,Caspase3,Caspase8,and TNF? genes in the normal tissues of Xuanwei females and non-Xuanwei females.(3)There was no significant difference in the expression of P100,Caspase3,Caspase8 and TNF? genes between normal and cancerous tissues in male and female patients.Conclusions:1.Nanosilica combined with benzo[a]pyrene can increase the migration and invasion of BEAS-2B cells compared to nanosilica and benzo[a]pyrene alone.2.The combination of nano-silica and benzo[a]pyrene was more effective than nano-silica and benzo[a]pyrene alone in the TNF?-TNFR signaling pathway.3.The effect of nano-silica combined with benzo[a]pyrene on TNF?-TNFR signaling pathway may be one of the causes of the high incidence of lung cancer in Xuanwei women.