title: Identification of novel NF-κB transcriptional targets in TNFα-treated HeLa and HepG2 cells.
abstract: Identification of target genes of NF-κB is critical for deeply understanding its biological functions. Here, we identified five novel NF-κB target genes. Firstly, we found that 20 NF-κB potential target genes (PTGs) identified by ChIP-Seq and Genechip assay were enriched into the KEGG term of Pathways in cancer, 16 of them were enriched into the KEGG pathways of small cell lung cancer, chronic myeloid leukemia, basal cell carcinoma, pancreatic cancer, and colorectal cancer. Among these PTGs, there are many documented NF-κB target genes. Therefore, NF-κB may play important role in cancer progression by transcriptionally regulating these genes. Apart from the known target genes, we also found some novel PTGs including CYCS, MITF, FZD1, FZD8, and PIAS1. We subsequently demonstrated whether NF-κB transcriptionally control the five PTGs. The ChIP-Seq assay revealed that NF-κB/p65 bound to these genes in TNFα-treated HeLa. The bioinformatic analysis indicated that the NF-κB binding regions (i.e., ChIP-Seq peaks) contained κB sites and NF-κB/RelA DNA-binding motif. The ChIP-qPCR assay also confirmed that NF-κB bound to these regions in both TNFα-treated HeLa and HepG2 cells. The reporter construct showed that NF-κB could regulate luciferase expression via its binding region. Finally, qPCR and Western blot assay demonstrated that NF-κB indeed regulated the expression of these genes in the TNFα-treated HeLa and HepG2 cells. In a word, CYCS, MITF, FZD1, FZD8, and PIAS1 were identified as bona fide NF-κB target genes. These findings provide more insights into the role of NF-κB in cancers.
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.|School of Life Sciences and Food Technology, Hanshan Normal University, Chaozhou, 521041, China.
authors.forename: Fei
authors.identifier: http://orcid.org/0000-0002-2970-0559
authors.initials: F
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.
authors.forename: Xinhui
authors.initials: X
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.
authors.forename: Danyang
authors.initials: D
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.
authors.forename: Jinke
authors.initials: J
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D047369:Chromatin Immunoprecipitation; D019295:Computational Biology; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D059372:Nucleotide Motifs / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D015203:Reproducibility of Results; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D016328:NF-kappa B; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha
keywords: CYCS; FZD1; FZD8; MITF; NF-κB target gene; PIAS1