Exploring The Mechanism Of The Effect Of Sphingomonasand Environmental Carcinogens On MUC16 Expression In Lung Epithelial Cells

Objective :To investigate the effect and pattern of gene expression of MUC16 and LINC00842 in lung microenvironment: Sphingomonas and environmental carcinogens on lung cancer and normal lung epithelial cells.Methods:1.Bioinformatic association analysis of LINC00842 and MUC16 in 84 lung cancer samples in Cosmic,NCI-GDC,Starbase,Cbioportal and other databases;q-PCR validation analysis and subpopulation characterization of LINC00842 and MUC16 lung cancer tissues.2.Establishment of lung epithelial cell-Sphingomonas-environmental carcinogen co-culture system: lung adenocarcinoma cell line H1734 and normal lung bronchial epithelial cells BEAS-2B were inoculated with Sphingomonas multiforme and Sphingomonas landracea box bacteria and incubated for 24 h,48h,72 h and 96 h,respectively;72h was selected as the optimal incubation time,and xylene,formaldehyde,benzo[a]pyrene,anthracene,ethidium bromide,cigarette filter extract,Ni SO4 and other environmental carcinogens.The expression of the target genes MUC16 and LINC00842 in lung epithelial cells in the co-culture system,as well as other high-frequency mutant genes EGFR,KRAS,TP53,TERT and MMP9 in lung cancer were detected by q RT-PCR.Results:1.Bioinformatic correlation analysis and q-PCR validation analysis of lung cancer tissues1.1 Bioinformatic correlation analysisMUC16 was ranked eighth in the Cosmic database of cancer tissues from lung cancer patients,and was located second in male lung cancer and third in female lung cancer in the NCI-GDC database,and showed a positive correlation trend with LINC00842.1.2 Lung cancer tissue q-PCR validation analysisBoth LINC00842 and MUC16 were highly expressed in lung cancer tissues,and both were positively correlated in cancer and paracancer.MUC16 tended to be overexpressed in the lung adenocarcinoma,female and young patient groups,while LINC00842 was overexpressed in a wider range of lung cancer patients,and was more highly expressed in males,stage III-IV patients and elderly patients.2.Changes in expression of key cellular genes in a cell-sphingomonas-environmental carcinogen co-culture model2.1 Lung adenocarcinoma cell-sphingomonas-environmental carcinogen co-culture systemLung adenocarcinoma cells H1734,in the formaldehyde,Bap,anthracene,EB and cigarette groups,Sphingomonas spp.alone or in cooperation,showed the ability to reduce the oncogenes EGFR,KRAS,TERT,MMP9 and MUC16,and the microecological effect was mainly in the form of balanced regulation of "low is up,high is down".2.2 Normal lung epithelial cells-Sphingomonas sp.-environmental carcinogens co-culture systemThe overall genetic fluctuations in BEAS-2B cells were all insignificant;the oncogenes EGFR,KRAS,TERT,MMP9,and TP53 were not significantly upregulated in BEAS-2B cells under environmental carcinogen(formaldehyde,Bap,anthracene,EB,and cigarette)exposure.Conclusion(s):1.LINC00842 and MUC16 were highly expressed in lung cancer tissues;in both lung cancer and paraneoplastic tissues: MUC16 and LINC00842 showed a significant positive correlation,and LINC00842 may be involved in regulating MUC16 expression.2.Normal lung epithelial cells BEAS-2B had better stability and tolerance than lung adenocarcinoma cells H1734 in a co-culture environment with Sphingomonas and environmental carcinogens;Sphingomonas had some protective effect,probably acting through upregulation of LINC00842 and MUC16 expression.