title: A functional pseudogene, NMRAL2P, is regulated by Nrf2 and serves as a coactivator of NQO1 in sulforaphane-treated colon cancer cells.
abstract: The anticancer agent sulforaphane (SFN) acts via multiple mechanisms to modulate gene expression, including the induction of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-dependent signaling and the inhibition of histone deacetylase activity. Transcriptomics studies were performed in SFN-treated human colon cancer cells and in nontransformed colonic epithelial cells in order to pursue new mechanistic leads.
  
  
  
  RNA-sequencing corroborated the expected changes in cancer-related pathways after SFN treatment. In addition to NAD(P)H quinone dehydrogenase 1 (NQO1) and other well-known Nrf2-dependent targets, SFN strongly induced the expression of Loc344887. This noncoding RNA was confirmed as a novel functional pseudogene for NmrA-like redox sensor 1, and was given the name NmrA-like redox sensor 2 pseudogene (NMRAL2P). Chromatin immunoprecipitation experiments corroborated the presence of Nrf2 interactions on the NMRAL2P genomic region, and interestingly, NMRAL2P also served as a coregulator of NQO1 in human colon cancer cells. Silencing of NMRAL2P via CRISPR/Cas9 genome-editing protected against SFN-mediated inhibition of cancer cell growth, colony formation, and migration.
  
  
  
  NMRAL2P is the first functional pseudogene to be identified both as a direct transcriptional target of Nrf2, and as a downstream regulator of Nrf2-dependent NQO1 induction. Further studies are warranted on NMRAL2P-Nrf2 crosstalk and the associated mechanisms of gene regulation.
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.
authors.forename: Gavin S
authors.initials: GS
authors.lastname: Johnson
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.affiliation: Linus Pauling Institute, Oregon State University, Corvallis, OR, USA.|School of Biological and Population Health Sciences, Oregon State University, Corvallis, OR, USA.
authors.forename: Laura M
authors.initials: LM
authors.lastname: Beaver
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.
authors.forename: W Mohaiza
authors.initials: WM
authors.lastname: Dashwood
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.
authors.forename: Deqiang
authors.initials: D
authors.lastname: Sun
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.
authors.forename: Praveen
authors.initials: P
authors.lastname: Rajendran
authors.affiliation: Linus Pauling Institute, Oregon State University, Corvallis, OR, USA.|Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Williams
authors.affiliation: Linus Pauling Institute, Oregon State University, Corvallis, OR, USA.|School of Biological and Population Health Sciences, Oregon State University, Corvallis, OR, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Ho
authors.affiliation: Center for Epigenetics & Disease Prevention, Institute of Biosciences & Technology, Texas A&M Health Science Center, Houston, TX, USA.|Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Department of Nutrition and Food Science, Texas A&M University, College Station, TX, USA.|Department of Molecular and Cellular Medicine, Texas A&M College of Medicine, College Station, TX, USA.
authors.forename: Roderick H
authors.initials: RH
authors.lastname: Dashwood
mesh_terms: D016588:Anticarcinogenic Agents / Q000494:pharmacology; D002471:Cell Transformation, Neoplastic; D003106:Colon / Q000378:metabolism; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D017879:Isothiocyanates / Q000494:pharmacology* / Q000627:therapeutic use; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D011544:Pseudogenes; D015398:Signal Transduction / Q000187:drug effects; D013454:Sulfoxides; D013861:Thiocyanates / Q000494:pharmacology
chemical_list: D016588:Anticarcinogenic Agents; D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2; D013454:Sulfoxides; D013861:Thiocyanates; D016660:NAD(P)H Dehydrogenase (Quinone); C016766:sulforaphane
keywords: CRISPR/Cas9; Colorectal cancer; Loc344887; noncoding RNA