Modulation of FAK, Akt, and p53 by stress release of the fibroblast-populated collagen matrix

The Journal of Surgical Research
Mark A CarlsonJon S Thompson

Abstract

Fibroblast survival in a three-dimensional collagen matrix is dependent in part upon the rigid anchorage of the matrix to tissue culture plastic. We hypothesized that focal adhesion kinase (FAK) and protein kinase B (Akt) would be activated and that the p53 level would be low in the rigidly anchored (attached) collagen matrix; loss of anchorage (detachment) was hypothesized to have the opposite effects. Human foreskin fibroblasts were cultured in attached bovine collagen matrices for 48 h before detachment as free-floating matrices. At various time points postrelease, matrix lysates were blotted for the proteins of interest, and the terminal deoxynucleotidyltransferase-mediated dUTP nick-end label assay was performed on both whole matrices and cytospin preparations. Irradiated monolayer fibroblasts were used as positive controls for the amount of p53 protein. Terminal deoxynucleotidyltransferase-mediated dUTP nick-end label positivity in attached versus detached matrices (at 24 h post detachment) was 0.7 +/- 03 versus 5.3 +/- 1.7% (P < 0.05, unpaired t test). FAK and Akt were phosphorylated (activated) in the attached matrix; there was a near complete of loss of both activated forms within 4 h of matrix detachment. Irradiated m...Continue Reading

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Citations

Feb 13, 2016·Toxicological Sciences : an Official Journal of the Society of Toxicology·Xingang WangJie Chao
Sep 25, 2015·The Journal of Investigative Dermatology·Jie ChaoMark A Carlson
Mar 23, 2005·Biochemical and Biophysical Research Communications·J WangC A McCulloch
Nov 26, 2010·Laboratory Investigation; a Journal of Technical Methods and Pathology·Jennifer E BondHoward Levinson
Jan 8, 2014·Wound Repair and Regeneration : Official Publication of the Wound Healing Society [and] the European Tissue Repair Society·Jie ChaoMark A Carlson
Jul 28, 2007·Molecular and Cellular Biochemistry·Mark A CarlsonJeremiah J Gums
Nov 27, 2015·American Journal of Physiology. Lung Cellular and Molecular Physiology·Haijun LiuJie Chao
Aug 19, 2006·The Journal of Investigative Dermatology·Mervi J TorisevaVeli-Matti Kähäri

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