title: Reactivation of p53 by a Cytoskeletal Sensor to Control the Balance Between DNA Damage and Tumor Dissemination.
abstract: BACKGROUND
  Abnormal cell migration and invasion underlie metastasis, and actomyosin contractility is a key regulator of tumor invasion. The links between cancer migratory behavior and DNA damage are poorly understood.
  
  
  METHODS
  Using 3D collagen systems to recapitulate melanoma extracellular matrix, we analyzed the relationship between the actomyosin cytoskeleton of migrating cells and DNA damage. We used multiple melanoma cell lines and microarray analysis to study changes in gene expression and in vivo intravital imaging (n = 7 mice per condition) to understand how DNA damage impacts invasive behavior. We used Protein Tissue Microarrays (n = 164 melanomas) and patient databases (n = 354 melanoma samples) to investigate the associations between markers of DNA damage and actomyosin cytoskeletal features. Data were analyzed with Student's and multiple t tests, Mann-Whitney's test, one-way analysis of variance, and Pearson correlation. All statistical tests were two-sided.
  
  
  RESULTS
  Melanoma cells with low levels of Rho-ROCK-driven actomyosin are subjected to oxidative stress-dependent DNA damage and ATM-mediated p53 protein stabilization. This results in a specific transcriptional signature enriched in DNA damage/oxidative stress responsive genes, including Tumor Protein p53 Inducible Protein 3 (TP53I3 or PIG3). PIG3, which functions in DNA damage repair, uses an unexpected catalytic mechanism to suppress Rho-ROCK activity and impair tumor invasion in vivo. This regulation was suppressed by antioxidants. Furthermore, PIG3 levels decreased while ROCK1/2 levels increased in human metastatic melanomas (ROCK1 vs PIG3; r = -0.2261, P < .0001; ROCK2 vs PIG3: r = -0.1381, P = .0093).
  
  
  CONCLUSIONS
  The results suggest using Rho-kinase inhibitors to reactivate the p53-PIG3 axis as a novel therapeutic strategy; we suggest that the use of antioxidants in melanoma should be very carefully evaluated.
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Cecilia
authors.initials: C
authors.lastname: Herraiz
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Fernando
authors.initials: F
authors.lastname: Calvo
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Pahini
authors.initials: P
authors.lastname: Pandya
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Gaia
authors.initials: G
authors.lastname: Cantelli
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Irene
authors.initials: I
authors.lastname: Rodriguez-Hernandez
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Jose L
authors.initials: JL
authors.lastname: Orgaz
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: NaRa
authors.initials: N
authors.lastname: Kang
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Tinghine
authors.initials: T
authors.lastname: Chu
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH).
authors.forename: Erik
authors.initials: E
authors.lastname: Sahai
authors.affiliation: Tumor Plasticity Laboratory, Randall Division of Cell and Molecular Biophysics, King's College London, London, UK (CH, PP, GC, IRH, JLO, NK, TC, VSM); Tumor Cell Biology Laboratory, Cancer Research UK London Research Institute, London, UK (FC, ES).Current affiliations: Tumor Microenvironment Team, Institute of Cancer Research, Chester Beatty Laboratories, London, UK (FC); Department of Biochemistry and Molecular Biology, School of Medicine, University of Murcia and IMIB-Arrixaca, Murcia, Spain (CH). victoria.sanz_moreno@kcl.ac.uk.
authors.forename: Victoria
authors.initials: V
authors.lastname: Sanz-Moreno
mesh_terms: D000205:Actomyosin; D000818:Animals; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000378:metabolism* / Q000473:pathology; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics*; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018384:Oxidative Stress; D040081:Protein Array Analysis; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D046888:Tissue Array Analysis; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D011518:Proto-Oncogene Proteins; D017382:Reactive Oxygen Species; C411296:TP53I3 protein, human; D016159:Tumor Suppressor Protein p53; D000205:Actomyosin; C515851:ROCK1 protein, human; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases