The Research About LncRNA NR046269 And Its Effect On The Expression Of Related Genes In Bronchial Epithelial Cells Infected With Talaromyces Marneyffei

Objective: To detect the expression of lnc RNA NR046269 and APOL1 in the early stage of bronchial epithelial cells infected with Talaromyces marneyffei.To Filter related lnc RNA and m RNA genes and discuss their correlations.Method:(1)using gene chip technology to detect Expression profile of lnc RNA and m RNA in BEAS-2B cells after spores of Talaromyces marneyffei infected with 4 hours.(2)Bronchial epithelial cells(BEAS-2B cells)were used as study subjects,The experiment was divided into control group and test group,The test group with TM spores infected BEAS-2B cells(MOI = 10-20:1)for 4 hours,Using q PCR to detect the reliability of the gene chip.(3)By bioinformatics,the correlation analysis between lnc RNA NR046269 and differential expression of m RNA in chip expression profile is analyzed,Screening lnc RNA NR046269-m RNA interrelation pairs.(4)BEAS-2B cells were transfected with lentivirus vector to construct a stable overexpression and low expression lnc RNA NR046269 BEAS-2B cell line.The experiment was divided into No-load negative control group,NR046269 overexpression group and NR046269 interference group.QPCR technique was used to detect the expression of lnc RNA NR046269 and m RNAs gen in negative control group? NR046269 overexpression group and NR046269 interference group.Western blotting assay was used to detect the expression of APOL1 protein in NR046269 negative control group.Result(1)The results of microarray analysis showed that 329 m RNAs and 519 lnc RNAs were differentially expressed.The results of q PCR were in consistent with the results of gene chip.(2)The results showed that lnc RNA NR046269 was significantly correlated with 154 m RNAs.(PCC0.97,P < 0.05),Pick out the part is related to the biological process of the infection stage and with lnc RNA NR046269 m RNAs significantly related part(APOL1?HMOX1?FOXO3A?LTC?ALPK2?LTB4?APOL1?MFSD12).(3)A stable overexpression and low expression lnc RNA NR046269 cell line was successfully constructed.The q PCR results showed that Compared with the negative control group,The expression of NR046269 was significantly higher in NR046269 overexpression group,and the expression of NR046269 in NR046269 interference group was significantly lower.At the same time,the gene expression of APOL1 in NR046269 overexpression group was significantly down-regulated.The gene expression of APOL1 in NR046269 interference group was upregulated significantly.Western blotting: compared with the negative control group,the expression of APOL protein was significantly decreased in the NR046269 overexpression group.The expression of APOL1 protein was up-regulated in NR046269 interference group.Conclusion(1)the results of microarray showed that lnc RNA and m RNA were differentially expressed after bronchial epithelial cells infected by Talaromyces marneffei spores.(2)A stable overexpression and low expression lnc RNA NR046269 of the BEAS-2B cell line was successfully constructed.The expression of APOL1 gene and protein in NR046269 overexpression group was significantly down-regulated,while that of APOL1 gene and protein expression in NR046269 interference group was significantly up-regulated.The results showed that: NR046269 had a negative regulatory effect on APOL1.