ABSTRACT
The aim of study was to identify the downstream target genes of CX43 by Human Transcriptome Array. Therefore, a gene microarray was generated which consists of CX43-overexpressed hepatocellular carcinoma (HCC) cells transfected with the constructed plasmid and negative controls to identify candidate genes. Integrated bioinformatic analysis was used to clarify biological functions of the identified genes, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction network, and survival analysis. The candidate genes were further validated by qRT-PCR in liver cancer tissues and CX43-silenced HCC cells. We have found the mRNA and protein levels of CX43 significantly upregulated in HCC cells transfected with CX43 constructed plasmid. We identified 928 differentially expressed genes including 394 upregulated and 534 downregulated genes, enriched in the cancer related functions and pathways by GO and KEGG pathway analysis. The protein-protein interaction network revealed 9 hub genes in this study. Statistical analysis indicated that upregulation of RALA and SRC was associated with poor prognosis in liver cancer. The differential expression of 2 candidate genes were further validated in HCC cells and tissues. In conclusion, protein-coding genes RALA and SRC could be target genes of CX43 and therapeutic targets for hepatocellular carcinoma.
Subject(s)
Carcinoma, Hepatocellular , Connexin 43 , Gene Expression Regulation, Neoplastic , Liver Neoplasms , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/physiopathology , Connexin 43/genetics , Connexin 43/metabolism , Gene Expression Profiling , Humans , Liver Neoplasms/genetics , Liver Neoplasms/physiopathologyABSTRACT
The pure rice (Oryza sativa, subsp. Indica, cv. Zhenshan 97B) chloroplast DNA was digested by restriction enzyme BamHI and the resulting fragments were ligated to the BamHI site of pBR322. One recombinant plasmid which contains a 19.3kb insertel DNA fragment was isolated from the clone bank and was named pOSB1. A physical map of the recombinant plasmid was constructed by cleavage with ten restriction endonucleases, and the gene rbcL was located on the pOSB1.
