Study Of The Inhibition Of The Migration And Invasion Of Hepatocellular Carcinoma Cells Caused By EDNRB Gene And Identification And Interaction Analysis Of Key Genes And MicroRNAs In Hepatocellular Carcinoma

PART 1 IDENTIFICATION AND INTERACTION ANALYSIS OF KEY GENES AND MICRORNAS IN HEPATOCELLULAR CARCINOMAObjective: To find the key genes and miRNAs as well as their potential molecular mechanisms in hepatocellular carcinoma(HCC).Methods: The microarray datasets GSE22058,GSE25097 and GSE57958,including mRNA and miRNA profiles,were downloaded from the GEO database and were analyzed using GEO2 R.Functional and pathway enrichment analyses were performed using the DAVID database,and the protein–protein interaction(PPI)network was constructed using the Cytoscape software.Finally,miRDB was applied to predict the targets of the differentially expressed miRNAs(DEMs).Results: A total of 115 differentially expressed genes(DEGs)were found in HCC,including 52 up-regulated genes and 63 down-regulated genes.The gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses from DAVID showed that up-regulated genes were significantly enriched in chromosome segregation and cell division,while the down-regulated genes were mainly involved in complement activation,protein activation cascades,carboxylic acid metabolic processes,oxoacid metabolic processes and the immune response.From the PPI network,the 18 nodes with the highest degree were screened as hub genes.Among them,ESR1 was found to have close interactions with FOXO1,CXCL12 and GNAO1.In addition,a total of 64 DEMs were identified,which included 58 up-regulated miRNAs and 6 down-regulated miRNAs.ESR1 was potentially targeted by five miRNAs,including hsa-mir-18 a and hsa-mir-221.Conclusion: The roles of DEMs like hsa-mir-221 in HCC through interactions with DEGs such as ESR1 and CXCL12 may provide new clues for the diagnosis and treatment of HCC patients.PART 2 STUDY OF THE INHIBITION OF THE MIGRATION AND INVASION OF HEPATOCELLULAR CARCINOMA CELLS CAUSED BY EDNRB GENEObjective: To investigate the influences of endothelin receptor B(EDNRB)gene on the migration and invasion abilities of human liver cancer cells.Methods: Real-time fluorescent quantitative PCR and western blot were used to detect the expression of EDNRB mRNAs and protein in liver cancer cell lines SMMC-7721 and Huh7 and normal liver cell lines LO2 and hepatocellular carcinoma(HCC)tissues and corresponding para-cancerous tissues.The lentivirus with EDNRB gene was infected into SMMC-7721 and Huh7 cell,and CCK-8 and Transwell assay were used to test the abilities of proliferation,migration and invasion of the infected liver cancer cells.Results: The expression of EDNRB mRNA in LO2 cell is higher than it in SMMC-7721 and Huh7,and its' expression in HCC tissues was lower than that in the corresponding para-cancerous tissues(p < 0.01).The expression level of EDNRB protein in HCC tissues was lower than that in the corresponding para-cancerous tissues,as the expression of EDNRB protein in liver cancer cell lines SMMC-7721 and Huh7 is lower than that in liver cell line LO2(p < 0.01).The overexpression of EDNRB gene in SMMC-7721 and Huh7 cell had no influences on the proliferation(both p > 0.05),otherwise it could inhibit the cells' migration and invasion(both p < 0.01).Conclusion: In HCC tissues,the expression of EDNRB gene is downregulated.The migration and invasion of liver cancer cells can be inhibited by EDNRB gene.