Construction And Transcriptomic Analysis Of SIRT6 Knockout Lung Adenocarcinoma A549 Cell Line

Objective:To explore the possible molecular mechanisms of the role of SIRT6 in lung adenocarcinoma A549 cells by constructing SIRT6 knockout lung adenocarcinoma A549 stable transgenic cell lines.Methods:(1)Based on the CRISPR/Cas9 target design principles,sg RNA(single-guide RNA)was designed through bioinformatics to construct the sg RNA-SIRT6 plasmid,packaged into lentivirus infected A549 cells,SIRT6 knockout A549 cells were screened by purinamycin and identified by concurrent genome sequencing.The knockout effect of SIRT6 protein was detected by Western blotting.(2)RNA Sequencing was used to detect the changes of differentially expressed genes(DEGs)and signaling pathways After SIRT6 gene knockout in human lung adenocarcinoma A549 cells.Results:(1)DNA sequencing results showed that the sg RNA-SIRT6 recombinant plasmid was successfully constructed,Western blotting results showed that SIRT6 was not expressed in cells transfected with sg RNA-SIRT6 recombinant plasmid.(2)A total of 379 DEGs were found by RNA sequencing,including 295 down-regulated genes and 84up-regulated genes(qvalue<0.05).GO enrichment analysis showed that differentially expressed genes(DEGs)were involved in bioadhesion,cell adhesion,and cell migration.KEGG enrichment analysis showed that DEGs were involved in several pathways,including chemical carcinogenesis,cytochrome P450 metabolism of drugs,cytochrome P450 metabolism of isobiotin,retinol metabolism,etc.Conclusion:The A549 cell line with SIRT6 knockout was successfully constructed.Transcriptional sequencing analysis revealed the DEGs regulated by SIRT6 in lung adenocarcinoma A549 cells and the related signaling pathways involved,providing scientific basis for subsequent studies on the biological function and molecular mechanism of SIRT6 in lung adenocarcinoma.