title: Cystatin C deficiency promotes epidermal dysplasia in K14-HPV16 transgenic mice.
abstract: BACKGROUND
  Cysteine protease cathepsins are important in extracellular matrix protein degradation, cell apoptosis, and angiogenesis. Mice lacking cathepsins are protected from tumor progression in several animal models, suggesting that the regulation of cathepsin activities controls the growth of various malignant tumors.
  
  
  RESULTS
  We tested the role of cathepsins using a mouse model of multistage epithelial carcinogenesis, in which the human keratin-14 promoter/enhancer drove the expression of human papillomavirus type 16 (HPV16) early region E6/E7 transgenes. During the progression of premalignant dysplasia, we observed increased expression of cysteine protease cathepsin S, but concomitantly reduced expression of cathepsin endogenous inhibitor cystatin C in the skin tissue extract. Absence of cystatin C in these transgenic mice resulted in more progression of dysplasia to carcinoma in situ on the face, ear, chest, and tail. Chest and ear skin extract real time PCR and immunoblot analysis, mouse serum sample ELISA, tissue immunohistological analysis, and tissue extract-mediated in vitro elastinolysis and collagenolysis assays demonstrated that cystatin C deficiency significantly increased cathepsin expression and activity. In skin from both the chest and ear, we found that the absence of cystatin C reduced epithelial cell apoptosis but increased proliferation. From the same tissue preparations, we detected significantly higher levels of pro-angiogenic laminin 5-derived γ2 peptides and concurrently increased neovascularization in cystatin C-deficient mice, compared to those from wild-type control mice.
  
  
  CONCLUSIONS
  Enhanced cathepsin expression and activity in cystatin C-deficient mice contributed to the progression of dysplasia by altering premalignant tissue epithelial proliferation, apoptosis, and neovascularization.
authors.affiliation: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Weifang
authors.initials: W
authors.lastname: Yu
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.forename: Michael A
authors.initials: MA
authors.lastname: Shi
authors.forename: Jianan
authors.initials: J
authors.lastname: Wang
authors.forename: Meixiang
authors.initials: M
authors.lastname: Xiang
authors.forename: Shiro
authors.initials: S
authors.lastname: Kitamoto
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.forename: Galina K
authors.initials: GK
authors.lastname: Sukhova
authors.forename: George F
authors.initials: GF
authors.lastname: Murphy
authors.forename: Gabriela
authors.initials: G
authors.lastname: Orasanu
authors.forename: Anders
authors.initials: A
authors.lastname: Grubb
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Shi
mesh_terms: D000818:Animals; D017209:Apoptosis; D002403:Cathepsins / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D055316:Cystatin C / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D018450:Disease Progression; D004817:Epidermis / Q000378:metabolism* / Q000473:pathology; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics; D006801:Humans; D006965:Hyperplasia; D015151:Immunoblotting; D053547:Keratin-14 / Q000235:genetics*; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D050725:Papillomavirus E7 Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: C525031:Cst3 protein, mouse; D055316:Cystatin C; C068651:E6 protein, Human papillomavirus type 16; D053547:Keratin-14; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D002403:Cathepsins; C028217:cathepsin S