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      Keratinocyte Motility Is Affected by UVA Radiation—A Comparison between Normal and Dysplastic Cells

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          Abstract

          UVA radiation induces multiple and complex changes in the skin, affecting epidermal cell behavior. This study reports the effects of UVA exposure on normal (HaCaT) and dysplastic (DOK) keratinocytes. The adherence, spreading and proliferation were investigated by time-lapse measurement of cell layer impedance on different matrix proteins. Prior to UVA exposure, the time required for adherence and spreading did not differ significantly for HaCaT and DOK cells, while spreading areas were larger for HaCaT cells. Under UVA exposure, HaCaT and DOK cells behavior differed in terms of movement and proliferation. The cells’ ability to cover the denuded surface and individual cell trajectories were recorded by time-lapse videomicroscopy, during wound healing experiments. Dysplastic keratinocytes showed more sensitivity to UVA, exhibiting transient deficiencies in directionality of movement and a delay in re-coating the denuded area. The actin cytoskeleton displayed a cortical organization immediately after irradiation, in both cell lines, similar to mock-irradiated cells. Post-irradiation, DOK cells displayed a better organization of stress fibers, persistent filopodia, and new, stronger focal contacts. In conclusion, after UVA exposure HaCaT and DOK cells showed a different behavior in terms of adherence, spreading, motility, proliferation, and actin cytoskeleton dynamics, with the dyplastic keratinocytes being more sensitive.

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          Most cited references41

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          FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly.

          Cell migration is a complex, highly regulated process that involves the continuous formation and disassembly of adhesions (adhesion turnover). Adhesion formation takes place at the leading edge of protrusions, whereas disassembly occurs both at the cell rear and at the base of protrusions. Despite the importance of these processes in migration, the mechanisms that regulate adhesion formation and disassembly remain largely unknown. Here we develop quantitative assays to measure the rate of incorporation of molecules into adhesions and the departure of these proteins from adhesions. Using these assays, we show that kinases and adaptor molecules, including focal adhesion kinase (FAK), Src, p130CAS, paxillin, extracellular signal-regulated kinase (ERK) and myosin light-chain kinase (MLCK) are critical for adhesion turnover at the cell front, a process central to migration.
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            Random versus directionally persistent cell migration.

            Directional migration is an important component of cell motility. Although the basic mechanisms of random cell movement are well characterized, no single model explains the complex regulation of directional migration. Multiple factors operate at each step of cell migration to stabilize lamellipodia and maintain directional migration. Factors such as the topography of the extracellular matrix, the cellular polarity machinery, receptor signalling, integrin trafficking, integrin co-receptors and actomyosin contraction converge on regulation of the Rho family of GTPases and the control of lamellipodial protrusions to promote directional migration.
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              Cell migration in tumors.

              Invasion of cancer cells into surrounding tissue and the vasculature is an initial step in tumor metastasis. This requires chemotactic migration of cancer cells, steered by protrusive activity of the cell membrane and its attachment to the extracellular matrix. Recent advances in intravital imaging and the development of an in vivo invasion assay have provided new insights into how cancer cell migration is regulated by elements of the local microenvironment, including the extracellular matrix architecture and other cell types found in primary tumors. These results, combined with new findings from in vitro studies, have led to new insights into the molecular mechanisms of cell protrusive activity and chemotactic migration during invasion and metastasis.
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                Author and article information

                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                07 June 2018
                June 2018
                : 19
                : 6
                : 1700
                Affiliations
                [1 ]Victor Babeș National Institute of Pathology, 99-101, Splaiul Independentei, 050096 Bucharest, Romania; cristina_niculite@ 123456yahoo.com (C.M.N.); aurs.bv@ 123456gmail.com (A.O.U.); lauraceafalan@ 123456yahoo.com (L.C.C.)
                [2 ]Department of Morphological Sciences, University of Medicine and Pharmacy Carol Davila, 8, Blvd. Eroilor Sanitari, 050474 Bucharest, Romania
                [3 ]Department of Anatomy, Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest, 91-95, Splaiul Independentei, 050095 Bucharest, Romania; nemar_59@ 123456yahoo.com
                [4 ]SC Biotehnos SA, 3-5, Gorunului Street, 075100 Otopeni, Romania; lolariu@ 123456biotehnos.com
                Author notes
                [* ]Correspondence: mircea.leabu@ 123456ivb.ro
                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0003-3668-0856
                Article
                ijms-19-01700
                10.3390/ijms19061700
                6032280
                29880745
                6563bed3-6ce5-44a5-9f8a-5fec580b9558
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 03 May 2018
                : 05 June 2018
                Categories
                Article

                Molecular biology
                cell motility,keratinocyte,dysplastic cell,wound healing,actin cytoskeleton,focal contacts

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