Verification of miR-33a-5p Target Gene Analysis and Its Effect on Proliferation of Human Lung Adenocarcinoma A549 Cells
Objective To investigate the effect of miR-33a-5p overexpression on the proliferation of human lung adenocarcinoma A549 cells and explore its possible mechanism.Methods A549 cells were transfected with miR-33a-5p mimics,CCK-8 and BrdU assay were applied to detect the cell proliferation.Transcriptome sequencing(RNA-seq)was used to detect transcriptome expression level.Targetscan(8.0),miRDB(2020),miRWalk(release_2022_01)and ENCORI(starbase,v2.0)were jointly used to screen potential target genes of miR-33a-5p,then intersected with differentially expressed genes of RNA-seq,and gene expression was veri-fied by RT-qPCR.Results RT-qPCR showed that miR-33a-5p was overexpressed in A549 cells(P<0.05).CCK-8 assay and BrdU assay showed that cell proliferation was considerably inhibited(Pall<0.05).The results of transcriptome sequencing showed 494 differential genes,266 of which were up-regulated and 228 down-reg-ulated.GO functional enrichment was mainly in the extracellular region,integral component of plasma mem-brane and receptor complexes,extracellular exosomes,extracellular matrix structural constituent,and calcium ion binding,etc.KEGG enrichment occurred mainly in the complement and coagulation cascades,insulin resis-tance,ABC transporters,and IL-17 signaling pathway,NOD-like receptor signaling pathway,NF-κB signaling pathway,etc.Target gene prediction analysis and RT-qPCR expression verification showed that MTHFD2,OS-BPL6,HA DHB,HMGCLL1,GUCY1A2 and DEPTOR were significantly decreased(P all<0.05).Conclusion miR-33a-5p may inhibit the proliferation of A549 cells by regulating the post-transcriptional levels of MTHFD2,OSBPL6,HA DHB,HMGCLL1,GUCY1A2,and DEPTOR genes.
万方数据
维普
