Title:Silencing the Transcription Factor FOSL1 in Hyperproliferative HaCaT Cells Makes Them Susceptible to IFN-γ
VOLUME: 16 ISSUE: 3
Author(s):Prelovskaya Anna, Mezentsev Alexandre*, Piruzian Eleonora, Soboleva Anna and Bruskin Sergey
Affiliation:Department of Functional Genomics, Federal Non-Profit Research Institution of Russian Academy of Sciences, NI Vavilov Institute of General Genetics, Moscow, Department of Functional Genomics of Federal Non-profit Research Institution of Russian Academy of Sciences, NI Vavilov Institute of General Genetics, P.O. Box: 119333, 3 Gubkina Street, Suite 303, Moscow, Russia, Department of Functional Genomics, Federal Non-Profit Research Institution of Russian Academy of Sciences, NI Vavilov Institute of General Genetics, Moscow, Department of Functional Genomics, Federal Non-Profit Research Institution of Russian Academy of Sciences, NI Vavilov Institute of General Genetics, Moscow, Department of Functional Genomics, Federal Non-Profit Research Institution of Russian Academy of Sciences, NI Vavilov Institute of General Genetics, Moscow
Keywords:Apoptosis, FOSL1, HaCaT, IFN-γ, IL17, psoriasis, TNF.
Abstract:Background: The immortalized human epidermal keratinocytes HaCaT often
serve as an experimental model of psoriasis. Previously, we reported that Th1 cytokines
TNF, IFN-γ and IL17 suppressed proliferation in cultured HaCaT cells. Our results also
suggested that at certain conditions, HaCaT cells could become susceptible to the named
cytokines.
Objective: The aim of this study was to analyze the consequences of FOSL1 silencing in
HaCaT cells and explore the role of the transcription factor FOSL1 in the above mentioned
antiproliferative effect.
Methods: Lentiviral transduction was used to knock FOSL1 down in HaCaT cells.
Changes in cell proliferation were assessed by analyzing the growth of transduced cells.
Changes in gene expression were examined by qPCR.
Results: We found that the treatment of FOSL1-deficient HaCaT cells with a combination
of TNF, IFN-γ and IL17 led to growth arrest and apoptosis. In turn, exposure of these cells
to the individual cytokines suggested that the presence of IFN-γ in the culture medium was
the primary cause of cell death. FOSL1-deficient cells exposed to IFN-γ exhibited a dramatic
shape change and intensive blebbing. Moreover, TNF and IL17 accelerated IFN-γ-
induced apoptosis. A comparative analysis of gene expression in control and FOSL1-
deficient HaCaT cells discovered that only TNF induced ten FOSL1 target genes previously
implicated in the pathogenesis of psoriasis, namely CCL2, FOSL1, HMOX1, IL8,
IL6, IVL, MGP, MMP1, MMP9, and PLAUR. In contrast, neither IFN-γ nor IL17 produced
similar changes in the expression profiles of the mentioned genes.
Conclusion: The obtained results suggest that the transcription factor FOSL1 protects
HaCaT cells from apoptosis triggered by IFN-γ. This study also reveals distinct roles of
Th1 cytokines in the regulation of FOSL1 target genes.
