Screening of Colon Cancer-associated lncRNA-miRNA Network by Bioinformatics Approach and its Functional Validation at Cellular Level
Objective To screen differentially expressed lncRNA and miRNA in colon cancer,and to explore their association with colon cancer prognosis and related biological functions.Meth-ods The lncRNA and miRNA expression profile data and clinical information of colon cancer pa-tients were collected from TCGA database,and R software was used to screen differentially ex-pressed lncRNAs and miRNAs;Single-factor Cox regression analysis was used to identify differ-entially expressed lncRNA and miRNA that were significantly associated with prognosis;the miRNet database was used to construct a prognostic-related lncRNA-miRNA network;mirPath v.3 software was used for functional enrichment analysis;R software was used to build a prog-nostic risk model and validate it.CCK-8 experiments,scratch experiments,flow cytometry,and qPCR were used to evaluate the effects of knocking down TSPEAR-AS2 on the proliferation,mi-gration,apoptosis and downstream miRNA expression of human colon cancer HCT116 cells.Re-sults 1823 differentially expressed lncRNAs and 524 differentially expressed miRNAs were iden-tified in colon cancer tissues;158 lncRNAs and 31 miRNAs were found to be associated with co-lon cancer prognosis;an lncRNA-miRNA network composed of 8 lncRNAs and 14 miRNAs was constructed;GO and KEGG enrichment analyses showed that the lncRNA-miRNA network was mainly involved in tumor-related biological functions and pathways;11 key genes that could effec-tively evaluate colon cancer prognostic characteristics were identified,the survival time of the high-risk group was significantly shorter than that of the low-risk group in the prognostic risk model constructed from these 11 key genes(P＜0.001),and the AUCs for predicting the survival of 1,3,and 5 years were 0.70,0.74,and 0.71,respectively.Knocking down TSPEAR-AS2 inhibited the proliferation,migration,and promoted apoptosis of HCT116 cells,and down-regulated the ex-pression of hsa-mir-4731-5p and hsa-mir-342-3p.Conclusion successful screening of colon cancer-associated lncRNA-miRNA network and the prognostic risk model constructed therefrom can has good predictive efficacy.TSPEAR-AS2 could be involved in regulating the proliferation,migra-tion,apoptosis,and downstream miRNA expression of colon cancer cells.
万方数据
维普
