Premium
miR‐93 downregulation of PTK6 attenuates TNFα/IFNγ mediated of colonic epithelial barrier dysfunction (1113.2)
Author(s) -
Haines Ricci,
Beard Richard,
Yuan Sarah,
Wu Mack
Publication year - 2014
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.28.1_supplement.1113.2
Subject(s) - gene knockdown , downregulation and upregulation , barrier function , cancer research , transfection , tight junction , biology , microbiology and biotechnology , immunology , apoptosis , cell culture , biochemistry , genetics , gene
The role of protein tyrosine kinase 6 (PTK6, also known as breast tumor kinase BRK) has been mainly studied in mechanisms underlying metastasis and cancer. However, it has also been shown that PTK6 null mice demonstrate particularly strong (greater than 10 fold increase in basal TER resistance measurement relative to PTK6+/+) colonic epithelial monolayers. To this end, we investigated the role of PTK6 in TNFα/IFNγ mediated disruption of colonic epithelial barrier function. Consistent with previous findings, PTK6 knockdown increased basal resistance in YAMC monolayers. In addition, we showed that PTK6 knockdown significantly attenuated TNFα/IFNγ mediated decrease in resistance of young adult mouse colonic epithelial cell (YAMC) monolayers. In order to determine mechanisms of endogenous regulation of PTK6, we performed microRNA prediction analysis on the 3’UTR of PTK6 and identified miR‐93 as being highly likely to target PTK6 in epithelial cells. We investigated the role of miR‐93 in regulating PTK6 in YAMCs and determined that transfection of miR‐93 similarly attenuated TNFα/IFNγ mediated decrease in TER. We also show by Western blot that miR‐93 significantly decreased PTK6 expression following stimulant challenge. Using immunocytochemistry and confocal microscopy, we further characterized the process by which PTK6 modulates epithelial barrier function in YAMCs. Our results indicate that miR‐93 targeting of PTK6 may serve as a novel therapeutic strategy in treating diseases involving inflammation driven hyperpermeability of the colonic epithelium. Grant Funding Source : Supported by VA Merit Review 5101BX000799, the NIH grants HL96640, HL61507 and HL70752