Keratinocyte apoptosis on type I collagen gel caused by lack of laminin 5/10/11 deposition and Akt signaling.
Cultured human foreskin keratinocytes (HFKs) adhere to and grow on nonfibrous collagen via integrin alpha2beta1. During incubation, the receptors used for adhesion are changed to integrins alpha3beta1 and alpha6beta4 and those receptors bind to laminin 5 which is deposited by keratinocytes themselves. In this report, we examined the behaviors of HFKs and transformed keratinocytes on collagen fibril gels. These cells adhered to and spread on collagen gels using integrin alpha2beta1. After several hours on collagen gels, however, cells became round and apoptosis occurred. The behavior of keratinocytes contrasted to that of fibroblasts that grew well even on collagen gel. At the point of apoptosis, integrins alpha2beta1 and alpha3beta1 were not found in the contact region of HFKs. Also, deposition of laminin 5 on collagen gel was not found despite the synthesis of mRNA for laminin 5 and laminin 10/11, while soluble laminin 5 protein is readily detectable. Phosporylation of Akt, which is known as a survival signal, was detected in HFKs cultured on coated collagen; however, the protein level and signals of Akt were dramatically decreased on collagen gel after 1 day of culture. These results indicate that collagen gel has different effects than nonfibrous collagen on HFKs and transformed keratinocytes and the interactions of integrin alpha3beta1 and laminin 5/10/11 are indispensable for maintenance of keratinocyte adhesion and survival.
Fujisaki
Hitomi
H
Nippi Research Institute of Biomatrix, Adachi-ku, Tokyo, 120-8601, Japan.
Hattori
Shunji
S
D017209:Apoptosis / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism*; D005782:Gels / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D038982:Integrin alpha2beta1 / Q000378:metabolism; D039201:Integrin alpha3beta1 / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D007797:Laminin / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D015398:Signal Transduction / Q000502:physiology; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D024042:Collagen Type I; D005782:Gels; D038982:Integrin alpha2beta1; D039201:Integrin alpha3beta1; D007797:Laminin; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D000097794:Kalinin; C494918:AKT1 protein, human