Biological effects of in vitro THz radiation exposure in human foetal fibroblasts.

In recent years, terahertz (THz) radiation has been widely used in a variety of applications: medical, security, telecommunications and military areas. However, few data are available on the biological effects of this type of electromagnetic radiation and the reported results, using different genetic or cellular assays, are quite discordant. This multidisciplinary study focuses on potential genotoxic and cytotoxic effects, evaluated by several end-points, associated with THz radiation. For this purpose, in vitro exposure of human foetal fibroblasts to low frequency THz radiation (0.1-0.15THz) was performed using a Compact Free Electron Laser. We did not observe an induction of DNA damage evaluated by Comet assay, phosphorylation of H2AX histone or telomere length modulation. In addiction, no induction of apoptosis or changes in pro-survival signalling proteins were detected. Moreover, our results indicated an increase in the total number of micronuclei and centromere positive micronuclei induction evaluated by CREST analysis, indicating that THz radiation could induce aneugenic rather than clastogenic effects, probably leading to chromosome loss. Furthermore, an increase of actin polymerization observed by ultrastructural analysis after THz irradiation, supports the hypothesis that an abnormal assembly of spindle proteins could lead to the observed chromosomal malsegregation.

[1]  Gerald J. Wilmink,et al.  Determination of death thresholds and identification of terahertz (THz)-specific gene expression signatures , 2010, BiOS.

[2]  J. Schlom,et al.  Local delivery of recombinant vaccinia virus encoding for neu counteracts growth of mammary tumors more efficiently than systemic delivery in neu transgenic mice , 2010, Cancer Immunology, Immunotherapy.

[3]  Thorsten Schrader,et al.  Terahertz Electromagnetic Fields (0.106 THz) Do Not Induce Manifest Genomic Damage In Vitro , 2012, PloS one.

[4]  A. Antoccia,et al.  Micronuclei, CREST-positive micronuclei and cell inactivation induced in Chinese hamster cells by radiation with different quality. , 2000, International journal of radiation biology.

[5]  H. Pasolli,et al.  Developmental roles for Srf, cortical cytoskeleton and cell shape in epidermal spindle orientation , 2011, Nature Cell Biology.

[6]  Bayrammurad Saparov,et al.  Corrigendum: Complex structures of different CaFe2As2 samples , 2015, Scientific Reports.

[7]  H.B. Wallace,et al.  Standoff Detection of Weapons and Contraband in the 100 GHz to 1 THz Region , 2007, IEEE Transactions on Antennas and Propagation.

[8]  D. Gisselsson.  Mitotic Instability in Cancer: Is There Method in the Madness? , 2005, Cell cycle.

[9]  T. Kleine-Ostmann,et al.  Spindle disturbances in human‐hamster hybrid (AL) cells induced by mobile communication frequency range signals , 2008, Bioelectromagnetics.

[10]  M. Kirsch‐Volders,et al.  Mitotic non-disjunction as a mechanism for in vitro aneuploidy induction by X-rays in primary human cells. , 1996, Mutagenesis.

[11]  W. Kwok,et al.  Emission of Coherent THz Radiation from Superconductors , 2007, Science.

[12]  E. Grosse,et al.  THz radiation from free electron lasers and its potential for cell and tissue studies. , 2002, Physics in medicine and biology.

[13]  Shuting Fan,et al.  The potential of terahertz imaging for cancer diagnosis: A review of investigations to date. , 2012, Quantitative imaging in medicine and surgery.

[14]  Hua Zhong,et al.  Terahertz Spectroscopy and Imaging for Defense and Security Applications , 2007, Proceedings of the IEEE.

[15]  M. Fenech.  The in vitro micronucleus technique. , 2000, Mutation research.

[16]  I. Spassovsky,et al.  High power THz sources and applications at ENEA-Frascati , 2014 .

[17]  P. Monaci,et al.  Gene-specific inhibition of breast carcinoma in BALB-neuT mice by active immunization with rat Neu or human ErbB receptors. , 2007, International journal of oncology.

[18]  A. Doria,et al.  THz Exposure of Whole Blood for the Study of Biological Effects on Human Lymphocytes , 2003, Journal of biological physics.

[19]  A. Antoccia,et al.  The role of telomere length modulation in delayed chromosome instability induced by ionizing radiation in human primary fibroblasts , 2013, Environmental and molecular mutagenesis.

[20]  C. Tanzarella,et al.  Immunofluorescent staining of kinetochores in micronuclei: a new assay for the detection of aneuploidy. , 1988, Mutation research.

[21]  Avraham Gover,et al.  Terahertz Radiation Increases Genomic Instability in Human Lymphocytes , 2008, Radiation research.

[22]  A. Antoccia,et al.  Telomere alterations and genomic instability in long-term cultures of normal human fibroblasts irradiated with X rays and protons. , 2011, Radiation protection dosimetry.

[23]  Andrew E. Pelling,et al.  Moesin Controls Cortical Rigidity, Cell Rounding, and Spindle Morphogenesis during Mitosis , 2008, Current Biology.

[24]  R. Bei,et al.  Resveratrol potentiates the in vitro and in vivo anti-tumoral effects of curcumin in head and neck carcinomas , 2014, Oncotarget.

[25]  Jessica E. Grundt,et al.  Development of a compact terahertz time-domain spectrometer for the measurement of the optical properties of biological tissues. , 2011, Journal of biomedical optics.

[26]  E. Blackburn,et al.  Structure and function of telomeres , 1991, Nature.

[27]  Olga Zeni,et al.  Tera-Hertz radiation in Biological Research, Investigations on Diagnostics and study on potential Genotoxic Effects , 2004 .

[28]  M Koch,et al.  Medical THz imaging: an investigation of histo-pathological samples. , 2002, Physics in medicine and biology.

[29]  Thorsten Schrader,et al.  Field Exposure and Dosimetry in the THz Frequency Range , 2014, IEEE Transactions on Terahertz Science and Technology.

[30]  D. Gisselsson,et al.  Telomere‐mediated mitotic disturbances in immortalized ovarian epithelial cells reproduce chromosomal losses and breakpoints from ovarian carcinoma , 2005, Genes, chromosomes & cancer.

[31]  Dustin G. Mixon,et al.  In vitro investigation of the biological effects associated with human dermal fibroblasts exposed to 2.52 THz radiation , 2011, Lasers in surgery and medicine.

[32]  E. Cosman,et al.  Ultrastructural Changes in Axons Following Exposure to Pulsed Radiofrequency Fields , 2009, Pain practice : the official journal of World Institute of Pain.

[33]  Thorsten Schrader,et al.  Terahertz Radiation at 0.380 THz and 2.520 THz Does Not Lead to DNA Damage in Skin Cells In Vitro , 2013, Radiation research.

[34]  Lyubov V. Titova,et al.  Intense THz pulses cause H2AX phosphorylation and activate DNA damage response in human skin tissue , 2013, Biomedical optics express.

[35]  A R Bishop,et al.  DNA Breathing Dynamics in the Presence of a Terahertz Field. , 2009, Physics letters. A.

[36]  C. Garrido,et al.  Targeting heat shock proteins in cancer. , 2013, Cancer letters.

[37]  Norihiko Sekine,et al.  At the Dawn of a New Era in Terahertz Technology , 2007, Proceedings of the IEEE.

[38]  T. Kleine-Ostmann,et al.  Terahertz Radiation Induces Spindle Disturbances in Human-Hamster Hybrid Cells , 2011, Radiation research.

[39]  P. Neurath,et al.  Chromosome Loss compared with Chromosome Size, Age and Sex of Subjects , 1970, Nature.

[40]  Thorsten Schrader,et al.  Spindle disturbances in human–hamster hybrid (AL) cells induced by the electrical component of the mobile communication frequency range signal , 2011, Bioelectromagnetics.

[41]  S. Perner,et al.  Quantifying telomere lengths of human individual chromosome arms by centromere-calibrated fluorescence in situ hybridization and digital imaging. , 2003, The American journal of pathology.

[42]  Joo-Hiuk Son,et al.  Nanoparticle-enabled terahertz imaging for cancer diagnosis. , 2009, Optics express.

[43]  Ronald A. DePinho,et al.  The age of cancer , 2000, Nature.

[44]  Jennifer L. Rohn,et al.  Changes in Ect2 Localization Couple Actomyosin-Dependent Cell Shape Changes to Mitotic Progression , 2012, Developmental cell.

[45]  M. Fenech,et al.  HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. , 2003, Mutation research.

[46]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[47]  E. Blackburn,et al.  Telomere states and cell fates , 2000, Nature.

[48]  N. Kolchanov,et al.  No DNA damage response and negligible genome-wide transcriptional changes in human embryonic stem cells exposed to terahertz radiation , 2015, Scientific Reports.

[49]  Dino Di Carlo,et al.  Increased Asymmetric and Multi-Daughter Cell Division in Mechanically Confined Microenvironments , 2012, PloS one.

[50]  Mark Lee,et al.  Searching for a Solid-State Terahertz Technology , 2007, Science.

[51]  A. Davies,et al.  INSTITUTE OF PHYSICS PUBLISHING PHYSICS IN MEDICINE AND BIOLOGY , 2001 .

[52]  Masayoshi Tonouchi,et al.  Cutting-edge terahertz technology , 2007 .

[53]  A. S. Ratushnyak,et al.  Effect of terahertz electromagnetic waves on neurons systems , 2008, 2008 IEEE Region 8 International Conference on Computational Technologies in Electrical and Electronics Engineering.

[54]  Gerald J. Wilmink,et al.  Invited Review Article: Current State of Research on Biological Effects of Terahertz Radiation , 2011 .

[55]  A. Sannino,et al.  CYTOGENETIC OBSERVATIONS IN HUMAN PERIPHERAL BLOOD LEUKOCYTES FOLLOWING IN VITRO EXPOSURE TO THz RADIATION: A PILOT STUDY , 2007, Health physics.

[56]  F. Galvano,et al.  Curcumin induces apoptosis in breast cancer cell lines and delays the growth of mammary tumors in neu transgenic mice. , 2013, Journal of biological regulators and homeostatic agents.

[57]  W. Bement,et al.  And the dead shall rise: actin and myosin return to the spindle. , 2011, Developmental cell.

[58]  F. Jankovics,et al.  Live imaging reveals that the Drosophila actin-binding ERM protein, moesin, co-localizes with the mitotic spindle. , 2009, European journal of cell biology.

[59]  Andrea Dimitracopoulos,et al.  Mitotic rounding alters cell geometry to ensure efficient bipolar spindle formation. , 2013, Developmental cell.

[60]  A. Antoccia,et al.  Telomere length in mammalian cells exposed to low- and high-LET radiations. , 2006, Radiation protection dosimetry.

[61]  J. Federici,et al.  THz imaging and sensing for security applications—explosives, weapons and drugs , 2005 .

[62]  E. Linfield,et al.  Terahertz pulse imaging in reflection geometry of human skin cancer and skin tissue. , 2002, Physics in medicine and biology.

[63]  P. H. Fitzgerald,et al.  Total aneuploidy and age-related sex chromosome aneuploidy in cultured lymphocytes of normal men and women , 1977, Human Genetics.

[64]  Michael S. Shur,et al.  Terahertz science and technology for military and security applications , 2007 .