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Decreased EGFR mRNA
expression in response to antipsoriatic drug dithranol in
vitro
Tewin Tencomnao1*,
Chalinee Ronpirin2, Anchalee Prasansuklab1
and Yong Poovorawan3
1Department
of Clinical Chemistry, Faculty of Allied Health Sciences,
Chulalongkorn University,
Bangkok 10330, Thailand.
2Department
of Preclinical Science, Faculty of Medicine, Thammasat
University, Pathumthani 12121, Thailand.
3Center
of Excellence in Clinical Virology, Department of Pediatrics,
Faculty of Medicine, Chulalongkorn University, Bangkok
10330, Thailand.
*Corresponding author. E-mail:
tewin.t@chula.ac.th. Phone: (662) 218-1081 ext. 313. Fax: (662) 218-1082.
Abbreviations: Bp,
Base pairs; DNase I, deoxyribonuclease; FBS,
fetal bovine serum; DMEM, dulbecco’s modified eagle’s
medium; EGF, epidermal growth factor; EGFR,
epidermal growth factor receptor; HB-EGF, heparin
binding EGF-like growth factor; PBS, phosphate
buffered saline; RT-PCR, reverse
transcriptase-polymerase chain reaction; TGF-α, transforming growth factor-α; VDRE,
vitamin D-responsive element.
Accepted 26
March, 2009 |
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Dithranol is enormously effective in the treatment of
psoriasis; however its molecular mode of action should be
further elucidated. Since epidermal growth factor receptor (EGFR)
is involved in the pathogenesis of psoriasis, the objective
of this study was to investigate the transcriptional effect
of dithranol on EGFR gene expression in the HaCaT
keratinocyte cell line, which is commonly employed as a
model system to study psoriasis including experiments
examining the effects of therapeutic drugs and cellular
regulators on keratinocytes. Cultured HaCaT cells were
treated with 0.1-0.5 µg/ml dithranol for 30 min. After 4 h,
total cellular RNA isolated from HaCaT cells was reverse
transcribed to cDNA which was subjected to polymerase chain
reaction (PCR) with specific primer pair for EGFR. We found
that dithranol treatment down-regulated the EGFR mRNA of
HaCaT cells in a concentration-dependent manner. Our result
was further substantiated using a
quantitative
real-time PCR approach. Taken together, the
dithranol-induced down-regulation of the EGFR in cultured
human keratinocytes might help to disclose the molecular
therapeutic action of the drug.
Key words:
Psoriasis, dithranol, EGFR, gene expression, human
keratinocytes. |
