Bioinformatics Analysis And Functional Study Of Circular RNA CircCSNK1G3 In Lung Adenocarcinoma

BackgroundCircular RNA(circRNA)is a new type of non-coding RNA that can regulate gene expression at the level of transcription or post-transcription.According to recent studies,circRNA is involved in the occurrence and development of various diseases,including cancers,but the role of circ RNA in lung adenocarcinoma is still unclear.In this study,bioinformatics was combined with experiments to screen and verify circRNAs involved in the pathogenesis of lung adenocarcinoma.MethodsFirst,the circRNA microarray was performed to screen the differentially expressed circRNAs in lung adenocarcinoma,to identify the suitable object.Using bioinformatics methods to predict the structure and function of circRNA,and a competitive endogenous RNA(ceRNA)network was constructed to predict the downstream miRNAs and mRNAs.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed based on the DAVID database to predict the functions and pathways.Subsequently,in order to verify the results of the circ RNA microarray and bioinformatics analysis,real-time fluorescence quantitative PCR(RT-qPCR)was used to evaluate the expression level of circ RNA.Si RNA was constructed and transfected into lung adenocarcinoma cells.MTT assay,Transwell assay and other cell experiments were conducted to observe the effects on proliferation,invasion and migration.Finally,animal experiment was used to observe its role in the development of lung adenocarcinoma.Results54 circRNAs were differentially expressed in lung adenocarcinoma.Among them,41 circRNAs were low expressed and 13 circRNAs were high expressed compared to normal tissues.By integrative analyzing the sequence,length and fold change of differential expression of circular RNA,we selected hsa?circ?0001522(called Circ CSNK1G3),which is highly expressed in tumors.Then,a total of 174 mRNA binding miRNAs and 39 circRNA binding miRNAs were found on the basis of miRNA ‘sponge' according to the database.GO result suggested that mRNAs were mainly rich in covalent chromatin modification and histone modification,which indicated that they might played roles in epigenetic regulation.In addition,KEGG showed that mRNAs were mainly rich in PI3K-Akt signaling pathway and cAMP signaling pathway.Then,RT-q PCR showed that the expression level of circCSNK1G3 in lung adenocarcinoma tissues and cells was significantly increased.We established an oncogenic role for circCSNK1G3 based on its aberrant expression in LUAD and associated enhancement of A549 and H1299 cell proliferation,migration and invasion.Animal experiments found that CircCSNK1G3 can promote the proliferation of lung adenocarcinoma in nude mice.ConclusionThe circRNA microarray and RT-qPCR showed that the expression of circCSN1G3 in lung adenocarcinoma tissues and cells increased significantly.The cell experiments and animal experiments indicated that circCSNK1G3 played a role in the development of lung adenocarcinoma.Combined with bioinformatics analysis,circ CSNK1G3 has the potential to promote the development of lung adenocarcinoma,and may be used as a potential therapeutic target for lung adenocarcinoma.