0007 -4888/11/1505?0632 © 2011 Springer Science+Business Media, Inc.
Effects of Expression of Transcriptional Factor AP-1
FOSL1 Gene on Psoriatic Process
V. V. Sobolev, A. D. Zolotorenko, A. G. Soboleva, A. M. Elkin*,
S. A. Il’ina*, D. N. Serov**, N. N. Potekaev**, S. B. Tkachenko**,
M. T. Minnibaev**, and A. L. Piruzyan*
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 150, No. 11, pp. 564-566, November, 2010
Original article submitted November 19, 2009
We performed quantitative analysis of FOSL1 gene expression in lesional psoriatic skin. The
expression of this gene in lesional psoriatic skin was signifi cantly increased compared to that
in unaffected areas. Enhanced FOSL1 expression signifi cantly correlated with high psoriasis
area and severity index (PASI). High level of FOSL1 gene expression was proposed to be a
marker of pathological process activity in psoriasis.
Key Words: psoriasis; gene expression; real time polymerase chain reaction; FOSL1
N. I. Vavilov Institute of General Genetics, Russian Academy of Sci-
ences, Moscow; *Center of Theoretical Problems of Physical and
Chemical Pharmacology, Russian Academy of Sciences, Moscow;
**I. M. Sechenov Moscow Medical Academy, Russia. Address for
correspondence: firstname.lastname@example.org. A. L. Piruzyan
Psoriasis is a skin disease, which is a complex geneti-
cally determined pathology and involves large groups
of interacting genes/proteins .
Transcriptome data processing showed that pso-
riasis determined by dysregulation of the genetic ap-
paratus functioning is associated with enhanced tran-
scription of numerous genes of transcriptional complex
AP-1 [1,2]. Transcriptional factor AP-1 is a group of
pairwise complexes formed by DNA-binding proteins
of the Jun, Fos, and ATF (activating transcription fac-
tor) families .
A key position in this complex is occupied by
FOSL1 gene, which controls numerous cell functions.
This gene is expressed in different types of cells with
different intensity [12,13].
The role FOSL1 as a proto-oncogene is now ac-
tively studied [4,5,8,10,11]. The main role of FOSL1
gene is determined by its overexpression in various
cell lines and clinical samples (epithelial cancer cells).
Expression of this gene was found in various types
of epithelial cells; enhanced FOSL1 expression was
also observed in fi broadenomas. Positive reaction for
FOSL1 was observed in some compartment of the
bowel in the apical layer of stratifi ed epithelium .
The aim of this study was to investigate changes
in FOSL1 expression in lesional psoriatic skin.
MATERIALS AND METHODS
Biopsy specimens of lesional psoriatic skin and unaf-
fected skin from patients with psoriasis (10 patients)
were taken under local anesthesia using dermatologi-
cal punch. The samples were immediately frozen in
liquid nitrogen, weighed, and grinded in a mortar,
avoiding sample thawing.
RNA isolation from biopsy samples was perfor-
med on Qiagen columns according to standard RNeasy
Mini Kit protocol.
Reverse transcription was carried out in 200 μl
PCR test tubes. To this end, 5× M-MLV RT buffer, M-
MLV reverse transcriptase (100 U, Promega), dNTP,
RNase inhibitor RNasin (20 U, Promega), random
hexanucleotide primers (Promega), and RNA up to a
fi nal concentration 100 ng/μl were added to the tube
and the mixture was thermostated for 1 h at 37oC.
Electrophoresis was performed in horizontal aga-
rose gel in TAE buffer (40 mM Tris-HСl, pH 8.0;
Bulletin of Experimental Biology and Medicine, Vol. 150, No. 5, March, 2011 GENETICS
1 mM EDTA; 20 mM CH3COOH); 2% agarose gels
Primers to FOSL1 mRNA were chosen using Vec-
tor NTI Advance 10 software. Annealing temperature
for each primer pair used in this study was chosen
empirically, taking the temperature calculated in Oli-
goCalculator software as the baseline. Human genomic
DNA and total human cDNA were used as a the matrix
during for choosing PCR conditions.
Real-time PCR was performed in 96-well optic plates
using fl uorescent-labeled oligonucleotide probes Taq-
Man®. The reaction was performed using 2.5× reaction
mixture with referent stain ROX (Syntol). Primers and
probes were synthesized by DNA-Synthesis company.
Amplifi cation was performed in a PCR-amplifi er
(Bio-Rad, iQ4) according to following program: de-
naturation at 95oC for 4 min (stage 1), denaturation at
94oC for 30 sec (stage 2), and annealing and elongation
at 60oC for 1 min (stage 3); stages 2-3 were repeated
50 times (stage 4). Genes and relevant primers with
probes are presented (Table 1).
Expression of the target genes was standardized by
the expression of housekeeping gene GAPDH.
For calculation of results the following para meters
were used: PCR effectiveness at least 95%; correlation
coeffi cient at least 0.99; slope -3.4±0.2.
Processing of PCR results was performed by
2—ΔΔCT method according to .
In previous bioinformatic investigations we used da-
tabase GEO DataSets (http://www.ncbi.nlm.nih.gov/
geo/), where the results of evaluation of gene expres-
sion levels on biochips are presented in electron tables.
MetaCore software (GenеGo Inc) was used as an
instrument for processing of tabulated data. Processing
of microarray data and the search psoriasis candidate
genes were performed using this software product. The
priority distribution of processes was carried out using
Metacore software, assuming that lower p-value cor-
responds to higher relevancy of the genes. The data of
bioinformatic analysis attested to the possible key role
of FOSL1 in the development of lesions and pathoge-
nesis of psoriasis .
The levels of FOSL1 expression in lesional psori-
atic skin were compared with those in unaffected skin
using Real Time PCR.
Patient state was assessed using psoriasis area
and severity index (PASI). Local PASI was estimated
for each part of the body. Integral PASI was equal
to the sum of local indices and could vary from 0 to
72 units. The maximum and minimum PASI values
in the given group were 11.4 and 1.8, respectively
The experiment showed that FOSL1 expression
in lesional psoriatic skin in virtually all patients was
2-fold higher than in unaffected skin. In patients 2,
4, 10, FOSL1 expression was almost 10-fold higher:
51.0321, 12.6699, and 22.3675, respectively (Fig. 1).
It should be noted that in patient with the highest PASI
values gene expression increased more than 10-fold;
high FOSL1 expression levels corresponded to higher
The data on the relationship between FOSL1
and psoriasis are contradictory. Some investigators
reported decreased expression of this gene in the skin
of patients with psoriasis , while in other studies,
changes in the expression of other members of AP-1
family, but not FOSL1 were observed . Appar-
ently, further investigations of FOSL1 as a gene par-
ticipating in psoriasis development are needed.
TABLE 1. Primers and Probes Used for Evaluation of Gene Expression Using Real-Time PCR
No.NameSequence 5’-3’PCR product length, b.p.s
Primer kit and probe were obtained from DNA-Synthesis200
Fig. 1. Changes in FOSL1 expression level in lesional psoriatic skin.
V. V. Sobolev, A. D. Zolotorenko, et al.
634 Download full-text
Generally, enhanced FOSL1 expression in all ex-
amined patients in our study attests to possible key
role of FOSL1 gene in psoriasis development.
The study was supported by Russian Foundation
for Basic Researches (grant No. 08-04-12136-ofi ) and
Presidium of Russian Academy of Sciences, program
“Fundamental Sciences for Medicine”.
TABLE 2. Clinical Parameters of Patients with Psoriasis
Regression of subjective signs:
itch, pain, tension
8 2.8 0.6
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