Transduction of beta3 integrin subunit cDNA confers on human keratinocytes the ability to adhere to gelatin.
alphavbeta3 is a multiligand integrin receptor that interacts with fibrinogen (FG), fibrin (FB), fibronectin (FN), vitronectin (VN), and denatured collagen. We previously reported that cultured normal human keratinocytes, like in vivo keratinocytes, do not express alphavbeta3 on the cell surface, and do not adhere to and migrate on FG and FB. Furthermore, we reported that human keratinocytes transduced with beta3 integrin subunit cDNA by a retrovirus-mediated transduction method express alphavbeta3 on the cell surface and adhere to FG, FB, FN, and VN significantly compared with beta-galactosidase (beta-gal) cDNA-transduced keratinocytes (control). In this study, we determined whether these beta3 integrin subunit cDNA-transduced keratinocytes or normal human keratinocytes adhere to denatured collagen (gelatin) using a 1 h cell adhesion assay. beta3 cDNA-transduced keratinocytes adhered to gelatin, whereas no significant adhesion was observed with the control cells (beta-gal cDNA-transduced keratinocytes and normal human keratinocytes). The adhesion to gelatin was inhibited by LM609, a monoclonal antibody to alphavbeta3, and RGD peptides but not by normal mouse IgG1 nor RGE peptides. Thus, transduction of beta3 integrin subunit cDNA confers on human keratinocytes the ability to adhere to denatured collagen (gelatin) as well as to FG, FB, VN, and FN. Otherwise, normal human keratinocytes do not adhere to gelatin. These data support the idea that beta3 cDNA-transduced human keratinocytes can be a good material for cultured epithelium to achieve better take rate with acute or chronic wounds, in which FG, FB, and denatured collagen are abundantly present.
Kubo
Miyoko
M
Department of Plastic and Reconstructive Surgery, Kawasaki Medical School, 577 Matsushima, Kurashiki City, Okayama, 701-0192, Japan. mkubo@med.kawasaki-m.ac.jp
Clark
Richard A F
RA
Katz
Anne B
AB
Taichman
Lorne B
LB
Jin
Zaishun
Z
Zhao
Ying
Y
Moriguchi
Takahiko
T
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics*; D005780:Gelatin / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000235:genetics / Q000276:immunology / Q000502:physiology; D039661:Integrin beta3 / Q000235:genetics* / Q000502:physiology; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D008297:Male; D009842:Oligopeptides / Q000276:immunology / Q000502:physiology; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D014161:Transduction, Genetic / Q000379:methods*
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D039302:Integrin alphaVbeta3; D039661:Integrin beta3; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid; D005780:Gelatin