[The establishment of the two dimensional gel electrophoresis (2-DE) map of hypertrophic scar epidermal cells and the display of the differentially expressed proteins].
OBJECTIVE
To establish and optimize the two dimensional gel electrophoresis (2-DE) map of epidermal cells separated from the hypertrophic scar tissues so as to explore the function of the non cultured epidermal cells during the course of the formation of hypertrophic scar.


METHODS
To separate the epidermal cells from the scar tissues, the scar epidermis was digested with Dispase II and trypsin. The total protein of the cells was then extracted and separated with 2-DE and visualized with silver stain. Spots detection and matching were performed with Melaine 3.0 gels analyzing software. The results were then compared with the normal epidermal cells' 2-DE map coming from the Danish Center for Human Genome Research' s 2-DE PAGE Databases.


RESULTS
Nearly more than 600 protein spots were identified in the final optimized 2-DE map. We found out 24 differentially expressed proteins by comparing the difference in composition, shape or density of all the spots. In the 24 proteins, there are 8 up-regulated ones, 9 down-regulated ones, 4 disappeared ones and 3 newly founded ones.


CONCLUSIONS
The method of digesting the epidermis with Dispase II and trypsin to separate the epidermal cells to establish the 2-DE map is feasible and it made the further study on hypertrophic scar proteomics possible. The 24 differentially expressed proteins revealed that epidermal cells might play a role in the formation of hypertrophic scar.
Zhang
Xi
X
Department of Plastic Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Gu
Shu-Zhong
SZ
Zhang
Lin-Xi
LX
Wu
Li-Hua
LH
Pan
Yang-Lin
YL
D002460:Cell Line; D002648:Child; D017439:Cicatrix, Hypertrophic / Q000378:metabolism* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004817:Epidermis / Q000378:metabolism*; D006801:Humans; D020543:Proteome / Q000378:metabolism*
D020543:Proteome