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Immortalized keratinocytes that overexpress H‐ras produce an invasive, randomized epithelium in organotypic culture
Author(s) -
Vaughan Melville B.,
Andrews Capri M.,
Wright Woodring E.,
Shay Jerry W.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.978.3
Subject(s) - involucrin , basement membrane , keratinocyte , keratin , epithelium , laminin , foreskin , fibroblast , chemistry , biology , pathology , cancer research , microbiology and biotechnology , in vitro , cell culture , medicine , biochemistry , extracellular matrix , genetics
Ras proteins affect proliferation and expression of collagen‐degrading enzymes, two important processes in cancer progression. Normal skin architecture is dependent both on the coordinated proliferation and stratification of keratinocytes, as well as the maintenance of a collagen‐rich basement membrane. Therefore we asked whether overexpression of H‐ras in skin keratinocytes would affect these parameters during the establishment and maintenance of an in vitro skin equivalent. Previously described immortalized foreskin keratinocytes were engineered to overexpress the H‐ras protein. Skin equivalents, composed of fibroblast‐contracted collagen gels overlaid with keratinocytes, were prepared and incubated for 3 weeks. Harvested tissues were processed and sectioned for histology and antibody staining. Antigens specific to proliferation (Ki67), differentiation (involucrin, keratin‐17, p63) and basement‐membrane formation (collagen IV, laminin‐5) were studied. Results showed that H‐ras keratinocytes produced an invasive, randomized epithelium most apparent in the lower strata. The superficial strata retained somewhat normal characteristics, however. Studies such as these may be helpful to understand cancer progression so that treatment or even prevention can be developed.