Abstract 1383: Impact of carbamylation and glycation of collagen type I on HT1080 cell migration
Authors: Said Georges , Guilbert Marie , Millerot-Serrurot Emilie , Van Gulick Laurence , Terryn Christine , Garnotel Roselyne , Jeannesson Pierre 
Published: June 2012

Abstract
Carbamylation and glycation are well known as nonenzymatic posttranslational modifications that occur throughout the lifespan of proteins in vivo. Due to its longevity, collagen type I, largely present in connective tissues, becomes a prominent target for such modifications. Protein carbamylation and glycation have been implicated in normal aging and in a number of pathologies like diabetes, renal diseases and cancer. In addition, recent studies suggest a possible link between increased cancer risk and diabetes or end-stage renal disease. Here, we investigate the impact of carbamylation and glycation of collagen on the proliferation and migration capacities of the highly invasive human fibrosarcoma cell line HT1080. For this purpose, collagen type I extracted from rat tail tendons was used for in vitro carbamylation with cyanate or glycation with ribose. The amount of carbamylation and glycation was evaluated respectively via HPLC and spectrofluorimetry, and only the maximum levels of carbamylated and glycated collagens were considered. Tumor cell behavior was then evaluated on coatings of native, carbamylated or glycated collagen. Our results show that the proliferation of HT1080 cells on modified collagens did not differ from that on native form after 1, 2 and 3 days of culture. Concerning tumor cell adhesion, the glycated collagen delayed the adhesion time of seeded cells whereas the carbamylated form had no effect. Using time-lapse videomicroscopy, the migration ability of HT1080 was studied by quantifying single cell speed. Only, the glycated collagen strongly inhibited cell speed migration. We next investigated the effect of these collagen modifications on the organization of actin and vinculin, two proteins involved in cell locomotion. On glycated collagen, cells revealed a dramatic loss of actin stress fibers where disorganized F-actin was principally localized at the rim of the cell. Disturbance of actin integrity was accompanied with a disorganization of vinculin that was also localized at the cell periphery. On the other hand, the cells on carbamylated collagen maintained a fully organized actin network. The impact on the focal adhesion kinase (FAK), which plays a crucial role in focal adhesion formation, was also evaluated. Only glycated collagen induced a significant inhibition of the expression level of FAK, whereas both collagen modifications provoke a differential inhibition of its phosphorylation state that is mainly taking place with glycation. In conclusion, only glycation is demonstrated as an important factor affecting tumor cell migration. This impact has to be certainly considered in order to better understand the link between diabetes, aging and cancer incidence.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1383. doi:10.1158/1538-7445.AM2011-1383