Cutaneous rat wounds express C49a, a novel gene with homology to the human melanoma differentiation associated gene, Mda-7

• Published in: Journal of cellular biochemistry 1999-07, Vol.74 (1), p.1-10
• Main Authors: Soo, Chia, Shaw, William W., Freymiller, Earl, Longaker, Michael T., Bertolami, Charles N., Chiu, Robert, Tieu, Andy, Ting, Kang
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cell Division - genetics; cell proliferation; Cloning, Molecular; Cytokines; differential display; DNA; fibroblasts; gene expression; Genes, Tumor Suppressor; Growth Substances - genetics; growth suppression; Humans; Intercellular Signaling Peptides and Proteins; Interleukins; Male; Melanoma - genetics; Molecular Sequence Data; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; Sequence Homology, Amino Acid; Skin - metabolism; Skin - pathology; wound edge; wound healing; Wound Healing - genetics; Wounds and Injuries - genetics; Wounds and Injuries - pathology
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/(SICI)1097-4644(19990701)74:1<1::AID-JCB1>3.0.CO;2-M

We have used DD‐PCR (differential display‐polymerase chain reaction) to identify new genes that are over‐ or underexpressed during wound repair. DD‐PCR performed on excisional wounds identified the expression of rat c49a. Cloning and sequence analysis of the rat c49a gene revealed high homology to a novel human melanoma differentiation associated gene, mda‐7. The human mda‐7 gene isolated from melanoma cell lines, has been linked with human melanoma differentiation, and growth suppression. Moreover, transfection of human mda‐7 constructs into human tumor cells suppresses the growth and colony formation of tumor cells from diverse origins. To confirm and relatively quantitate expression of rat c49a gene during repair, specific primer, reduced cycle RT‐PCR (reverse transcription‐PCR) was performed. RT‐PCR showed an ∼9 to 12‐fold elevation of rat c49a mRNA at 12 h to 5 days above nonwounded controls that gradually decreased to ∼1.5 to 3‐fold by day 14. Cloning and sequence analysis of the entire 1200 base pair c49a gene product showed 78% nucleotide homology to human mda‐7. Immunohistochemistry studies localized rat C49A expression primarily to fibroblast‐like cells at the wound edge and base. The marked up‐regulation of rat c49a transcripts during the inflammatory and early granulation tissue phases of wound repair where cellular processes such as re‐epithelialization, angiogenesis, and fibroplasia predominate—suggest that c49a is associated with proliferation of fibroblasts in wound healing. J. Cell. Biochem. 74:1–10, 1999. © 1999 Wiley‐Liss, Inc.