H-ras expression in immortalized keratinocytes produces an invasive epithelium in cultured skin equivalents.

PloS One
Melville B VaughanJerry W Shay

Abstract

Ras proteins affect both 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. In the present studies we sought to determine whether expression of H-ras in skin keratinocytes would affect these parameters during the establishment and maintenance of an in vitro skin equivalent. Previously described cdk4 and hTERT immortalized foreskin keratinocytes were engineered to express ectopically introduced H-ras. Skin equivalents, composed of normal fibroblast-contracted collagen gels overlaid with keratinocytes (immortal or immortal expressing H-ras), were prepared and incubated for 3 weeks. Harvested tissues were processed and sectioned for histology and antibody staining. Antigens specific to differentiation (involucrin, keratin-14, p63), basement-membrane formation (collagen IV, laminin-5), and epithelial to mesenchymal transition (EMT; e-cadherin, vimentin) were studied. Results showed that H-ras keratinocytes produced an invasive, disorganized epithelium most apparent in the lower strata while immortalize...Continue Reading

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Citations

Jun 12, 2014·BMC Cell Biology·Candice A M SauderSusan E Clare
Jul 30, 2010·American Journal of Physiology. Renal Physiology·Toru SakairiJeffrey B Kopp
Nov 11, 2010·Cancer Research·Emilios GemenetzidisMuy-Teck Teh
Mar 30, 2011·PloS One·Chiou-Mei LeeYu-Sun Chang
May 10, 2011·Journal of Dental Research·E L SchellerP C Edwards
Dec 15, 2015·Biotechnology Journal·Mohammad M MovahedniaTong Cao
Sep 17, 2010·Biology of the Cell·Timothy J UnderwoodJeremy P Blaydes
Jun 21, 2013·PloS One·Hans K GhayeeJerry W Shay
Apr 14, 2016·International Journal of Cancer. Journal International Du Cancer·Neus CantariñoMarcus Buschbeck
Nov 5, 2016·Cancer Research·Jun-Han LeeJoseph H Jeong

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Methods Mentioned

BETA
genetic modifications
xenografting

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