Regulation of human Cripto-1 expression by nuclear receptors and DNA promoter methylation in human embryonal and breast cancer cells

J Cell Physiol. 2013 Jun;228(6):1174-88. doi: 10.1002/jcp.24271.

Abstract

Human Cripto-1 (CR-1) plays an important role in regulating embryonic development while also regulating various stages of tumor progression. However, mechanisms that regulate CR-1 expression during embryogenesis and tumorigenesis are still not well defined. In the present study, we investigated the effects of two nuclear receptors, liver receptor homolog (LRH)-1 and germ cell nuclear factor receptor (GCNF) and epigenetic modifications on CR-1 gene expression in NTERA-2 human embryonal carcinoma cells and in breast cancer cells. CR-1 expression in NTERA-2 cells was positively regulated by LRH-1 through direct binding to a DR0 element within the CR-1 promoter, while GCNF strongly suppressed CR-1 expression in these cells. In addition, the CR-1 promoter was unmethylated in NTERA-2 cells, while T47D, ZR75-1, and MCF7 breast cancer cells showed high levels of CR-1 promoter methylation and low CR-1 mRNA and protein expression. Treatment of breast cancer cells with a demethylating agent and histone deacetylase inhibitors reduced methylation of the CR-1 promoter and reactivated CR-1 mRNA and protein expression in these cells, promoting migration and invasion of breast cancer cells. Analysis of a breast cancer tissue array revealed that CR-1 was highly expressed in the majority of human breast tumors, suggesting that CR-1 expression in breast cancer cell lines might not be representative of in vivo expression. Collectively, these findings offer some insight into the transcriptional regulation of CR-1 gene expression and its critical role in the pathogenesis of human cancer.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azacitidine / analogs & derivatives
  • Azacitidine / pharmacology
  • Binding Sites
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology
  • Carcinoma, Ductal, Breast / genetics
  • Carcinoma, Ductal, Breast / metabolism*
  • Carcinoma, Ductal, Breast / pathology
  • Carcinoma, Embryonal / genetics
  • Carcinoma, Embryonal / metabolism*
  • Carcinoma, Embryonal / pathology
  • Cell Movement
  • DNA Methylation* / drug effects
  • DNA Modification Methylases / antagonists & inhibitors
  • DNA Modification Methylases / metabolism
  • Decitabine
  • Dose-Response Relationship, Drug
  • Embryonal Carcinoma Stem Cells / metabolism
  • Embryonal Carcinoma Stem Cells / pathology
  • Female
  • GPI-Linked Proteins / genetics
  • GPI-Linked Proteins / metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Neoplastic
  • Genes, Reporter
  • Histone Deacetylase Inhibitors / pharmacology
  • Humans
  • Hydroxamic Acids / pharmacology
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • MCF-7 Cells
  • Neoplasm Invasiveness
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Nuclear Receptor Subfamily 6, Group A, Member 1 / genetics
  • Nuclear Receptor Subfamily 6, Group A, Member 1 / metabolism*
  • Promoter Regions, Genetic*
  • RNA Interference
  • RNA, Messenger / metabolism
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Time Factors
  • Tissue Array Analysis
  • Transcription, Genetic
  • Transfection
  • Tretinoin / pharmacology
  • Valproic Acid / pharmacology

Substances

  • GPI-Linked Proteins
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Intercellular Signaling Peptides and Proteins
  • NR5A2 protein, human
  • NR6A1 protein, human
  • Neoplasm Proteins
  • Nuclear Receptor Subfamily 6, Group A, Member 1
  • RNA, Messenger
  • Receptors, Cytoplasmic and Nuclear
  • TDGF1 protein, human
  • trichostatin A
  • Tretinoin
  • Valproic Acid
  • Decitabine
  • Luciferases
  • DNA Modification Methylases
  • Azacitidine