Predictors of the scleroderma phenotype in fibroblasts from systemic sclerosis patients.
BACKGROUND
Fibroblasts from skin lesions generally exhibit the scleroderma phenotype in the early stage of in vitro culture, and act as one of the most important samples to investigate systemic sclerosis. However, not all cell lines from patients show the scleroderma phenotype, and little prospective evidence can be relied on to predict the phenotype of fibroblasts from systemic sclerosis patients.


OBJECTIVE
This study aims to find the predictive factors related to fibroblast phenotypes.


METHODS
Fibroblast lines from 15 patients with systemic sclerosis were established through explant culture, with their scleroderma phenotypes being determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) of procollagen transcription. Logistic regression analysis was used to analyse the clinical data of these patients. Haematoxylin-eosin staining was performed to observe histopathological differences. Expression of methylation-related factors was detected by real-time reverse transcription-polymerase chain reaction.


RESULTS
Eight of the 15 fibroblast cell lines from patients with systemic sclerosis exhibited the scleroderma phenotype. Logistic regression analysis yielded an equation (Y = -9.718 + 2.525X1) in which X1 significantly represents the Valentini Disease Activity Index. Histopathology results demonstrated that the scleroderma phenotype was positively correlated with leucocyte infiltration. Further laboratory observations showed increased expression of Dnmt1 in the positive phenotype fibroblasts and a positive correlation between elevations of Dnmt1 and the maintenance period of the scleroderma phenotype.


CONCLUSIONS
The results of this study suggest that the Valentini Disease Activity Index and expression of Dnmt1 may act as indicators of the scleroderma phenotype in fibroblasts. Additionally, these data also imply that epigenetic factors, such as Dnmt1, may be involved in the maintenance of the scleroderma phenotype. Our analyses may be beneficial to the research in systemic sclerosis.
Qi
Q
Q
Department of Dermatology, Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Guo
Q
Q
Tan
G
G
Mao
Y
Y
Tang
H
H
Zhou
C
C
Zeng
F
F
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D005347:Fibroblasts / Q000473:pathology; D006801:Humans; D016015:Logistic Models; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012595:Scleroderma, Systemic / Q000235:genetics / Q000473:pathology*; D014158:Transcription, Genetic
D017931:DNA Primers