Neurofibromatosis type 1 tumour suppressor gene expression is deficient in psoriatic skin in vivo and in vitro: a potential link to increased Ras activity.
BACKGROUND
Neurofibromatosis type 1 (NF1) protein (neurofibromin) accelerates the inactivation of Ras-GTP in various cell types. Somatic mutations of the NF1 gene may lead to malignant transformation and uncontrolled proliferation. We have previously shown that NF1 protein expression is downregulated in psoriasis in vivo.


OBJECTIVE
To study the functional expression and distribution of NF1 mRNA and protein in vivo and in psoriatic and normal keratinocyte cultures.


METHODS
Immunohistochemistry and in situ hybridization were used to study NF1 gene and protein expression in psoriasis in vivo. Furthermore, Northern and in situ hybridizations, immunoblot and localization analyses were utilized to study NF1 mRNA and protein in vitro in keratinocyte cultures.


RESULTS
NF1 tumour suppressor gene expression was reduced in lesional psoriatic skin compared with perilesional and normal skin in vivo. The in vitro results showed that the levels of NF1 mRNA and protein were reduced in cultured psoriatic keratinocytes during cellular differentiation even after multiple passaging of the cells. Moreover, cultured nonlesional psoriatic keratinocytes were almost equally defective as lesional cells with respect to NF1 expression.


CONCLUSIONS
Our findings demonstrate that psoriatic keratinocytes maintain an altered phenotype and gene expression profile even when isolated from interaction with lymphocytes and fibroblasts, which are known to increase proliferation of keratinocytes. As NF1 protein is regarded as a Ras proto-oncogene regulator, the aberrant expression and distribution of NF1 protein and mRNA found in the present study may be causative to the previously described increased activation of Ras in psoriatic lesions, and relate to altered cellular behaviour.
Karvonen
S-L
SL
Department of Anatomy and Cell Biology, University of Oulu, PB 5000, Aapistie 7, 90014 Oulu, Finland.
Koivunen
J
J
Nissinen
M
M
Ylä-Outinen
H
H
Björkstrand
A-S
AS
Peltonen
J
J
D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D015870:Gene Expression; D016514:Genes, Neurofibromatosis 1; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes; D025542:Neurofibromin 1 / Q000032:analysis* / Q000235:genetics; D000090063:Proto-Oncogene Mas; D011565:Psoriasis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
C000715470:MAS1 protein, human; D025542:Neurofibromin 1; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger