Gene expression of early hypertrophic scar tissue screened by means of cDNA microarrays.
BACKGROUND
Hypertrophic scar is an excessive healing response that often follows thermal injury. The most outstanding morphologic change is the overdeposition of collagen, which is caused by imbalance between synthesis and metabolism of collagen. Previous studies also found that transforming growth factor-beta was the key factor controlling scar formation. However, neither anti-transforming growth factor-beta nor other methods could completely control scar formation and contraction. This fact suggests a multifactorial cause. Fortunately, cDNA microarray throws light on the general alteration at the gene level, and thus could allow us to find some new clues for understanding scar formation and contraction.


METHODS
In this article, we report the results obtained from the scanning of gene expression of hypertrophic scar by means of cDNA microarray. Five cases of early human postburn hypertrophic scars were selected. Total tissue RNA was extracted from each hypertrophic scar sample and the corresponding uninjured region skin tissue; mRNA was further purified by Oligotex and then was reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray containing 4,096 genes on a type of chemical material-coated glass slide. After high-stringent washing, the hybridized slides were scanned for fluorescent signal detection. Then, the expression and distribution of cytoskeletal genes such as alpha-smooth muscle actin (alpha-SMA) gene; fibroblast tropomyosin TM30(pl) gene; vimentin gene; profilin gene; and BM40 gene of hypertrophic scar at 3, 6, 9, and 12 months age were further quantitatively studied by in situ hybridization or immunohistochemistry.


RESULTS
Our data indicated that there were 94 genes overexpressed and 3 genes down-regulated in early postburn hypertrophic scar. These altered genes were related to proto-oncogenes, apoptosis, immune regulatory genes, cytoskeletal elements, metabolism, and so forth. We also found that the detected cytoskeletal gene expression was much more intense at all time points than the control group. Consistent with clinical observation, cytoskeletal genes reached a peak at an early stage and gradually decreased.


CONCLUSIONS
Our study implied that multiple genes are involved in scar formation and contraction. Interferon is an autosecreted cytokine that might be responsible for self-control of overgrowth of cells in wounds. The early period of hypertrophic scar formation might be a good time for preventing overgrowth and contraction of hypertrophic scar by gene therapy.
Wu
Jun
J
Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing, People's Republic of China. junwupro@public.cta.cq.cn
Ma
Bing
B
Yi
Shaoxuan
S
Wang
Zhenxiang
Z
He
Weifeng
W
Luo
Gaoxing
G
Chen
Xiwei
X
Wang
Xihua
X
Chen
Alex
A
Barisoni
Dino
D
D000199:Actins / Q000235:genetics; D000328:Adult; D002056:Burns; D016022:Case-Control Studies; D002675:Child, Preschool; D017439:Cicatrix, Hypertrophic / Q000235:genetics* / Q000473:pathology; D003598:Cytoskeletal Proteins / Q000235:genetics; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D009124:Muscle Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis
D000199:Actins; D003598:Cytoskeletal Proteins; D009124:Muscle Proteins; C470306:Smooth muscle protein, human