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Characterization of a transcriptional repressor region of the human N‐cadherin gene
Author(s) -
Marley Kate,
Flitcroft Crysta,
Workman Aspen,
Madden Scott
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.495.17
Subject(s) - cadherin , repressor , transfection , biology , microbiology and biotechnology , reporter gene , gene , promoter , hela , motility , endogeny , cell culture , epigenetics , regulation of gene expression , gene expression , cell , genetics , biochemistry
Cadherins are calcium‐dependent transmembrane cell adhesion proteins. N‐cadherin can become misexpressed in epithelial tumors and is linked to increased motility and invasiveness. Loss of N‐cadherin expression in rat kidney cells is associated with degradation of kidney integrity and the N‐cadherin promoter becomes methylated in these cells. The objective of this study was to characterize a putative repressor region of the human N‐cadherin gene. N‐cadherin promoter luciferase reporter constructs were transiently transfected into HeLa cells and MDA‐MB‐435 cells, which both express N‐cadherin endogenously. Expression levels from reporters were highest from the +10bp to ‐462bp fragment and lowest from the +10bp to ‐1975bp fragment. Deletion reporters from ‐462bp to ‐1975bp are narrowing the region of focus for a relevant repressor element. A431 cells and BT‐20 cells do not express endogenous N‐cadherin and A431 also did not express any N‐cadherin reporters. However, BT‐20 cells had reporter expression similar to HeLa and MDA‐MB‐435 cells, suggesting there may be epigenetic modifications of the endogenous N‐cadherin promoter. Examination of the methylation status of the human N‐cadherin gene in BT‐20 cells in underway. This publication was made possible by the NIH Grant Number P20 RR16469 from the INBRE Program of the National Center for Research Resources.