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Development in Night Shift Workers  \nSilvia Vivarelli 1, *, Giovanna Spatari 1, Chiara Costa 2, Federica Giambò 1 and Concettina Fenga 1  \nCitation: Vivarelli, S.; Spatari, G.; Costa, C.; Giambò, F.; Fenga, C. Computational Analyses Reveal Deregulated Clock Genes Associated with Breast Cancer Development in Night Shift Workers. Int. J. Mol. Sci. 2024, 25, 8659. [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.3390/ijms25168659](10.3390/ijms25168659)  \nAcademic Editor: Marcella Mauro  \nReceived: 15 July 2024  \nRevised: 2 August 2024  \nAccepted: 7 August 2024  \nPublished: 8 August 2024  \nCopyright: © 2024 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 ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) .  \n1 Department of Biomedical and Dental Sciences, Morphological and Functional Imaging, Section of Occupational Medicine, University of Messina, 78712 Messina, Italy; giovanna.spatari@unime.it (G.S.); [federicagiambo@gmail.com](federicagiambo@gmail.com) (F.G.); [cfenga@unime.it](cfenga@unime.it) (C.F.)  \n2 Department of Clinical and Experimental Medicine, University of Messina, 78712 Messina, Italy; [chiara.costa@unime.it](chiara.costa@unime.it)  \n* Correspondence: [silvia.vivarelli@unime.it](silvia.vivarelli@unime.it)  \nAbstract: Breast cancer (BC) is the leading cause of cancer death among women worldwide. Women employed in shift jobs face heightened BC risk due to prolonged exposure to night shift work (NSW), classified as potentially carcinogenic by the International Agency for Research on Cancer (IARC) . This risk is linked to disruptions in circadian rhythms governed by clock genes at the cellular level. However, the molecular mechanisms are unclear. This study aimed to assess clock genes as potential BC biomarkers among women exposed to long-term NSW. Clock gene expression was analysed in paired BC and normal breast tissues within Nurses’ Health Studies I and II GEO datasets. Validation was performed on additional gene expression datasets from healthy night shift workers and women with varying BC susceptibility, as well as single-cell sequencing datasets. Post-transcriptional regulators of clock genes were identified through miRNA analyses. Significant alterations in clock gene expression in BC compared to normal tissues were found. BHLHE40, CIART, CLOCK, PDPK1, and TIMELESS were over-expressed, while HLF, NFIL3, NPAS3, PER1, PER3, SIM1, and TEF were under-expressed. The downregulation of PER1 and TEF and upregulation of CLOCK correlated with increased BC risk in healthy women. Also, twenty-six miRNAs, including miR-10a, miR-21, miR-107, and miR-34, were identified as potential post-transcriptional regulators influenced by NSW. In conclusion, a panel of clock genes and circadian miRNAs are suggested as BC susceptibility biomarkers among night shift workers, supporting implications for risk stratification and early detection strategies.  \nKeywords: night shift work; breast cancer; clock genes; micro-RNAs; circadian rhythms; biomarkers; computational study  \n1. Introduction  \nBreast cancer (BC) represents the most frequently diagnosed cancer among women worldwide. According to the Global Cancer Observatory, in 2022, 2.3 million women were diagnosed with BC, and 670,000 deaths have been counted globally [1] . In Europe, for the year 2024, it has been predicted that there will be the highest absolute number of deaths (89,300) in women [2] . Intriguingly, compared to less developed countries, in more developed ones there is a higher survival rate attributed to earlier diagnosis, better preventive measures, and improved access to treatment [3] . The aetiology of BC is multifaceted as it in","cbCaieWu2pMq1ImN","https://ap.wps.com/l/cbCaieWu2pMq1ImN","pdf",5335633,18,"English","# Abstract\n# Introduction","[{\"question\":\"What is the main research goal of the study?\",\"answer\":\"To assess clock genes as potential breast cancer biomarkers among women exposed to long-term night shift work.\"},{\"question\":\"Which data types were used to analyze clock gene expression?\",\"answer\":\"Paired breast cancer and normal tissues from Nurses’ Health Studies I and II GEO datasets, with validation using additional gene expression datasets, varying susceptibility groups, and single-cell sequencing datasets.\"},{\"question\":\"How were post-transcriptional regulators of clock genes identified?\",\"answer\":\"Through miRNA analyses to detect miRNAs that potentially regulate clock gene expression in the context of night shift work exposure.\"}]","Computational Analyses Reveal Deregulated Clock Genes Associated with Breast Cancer Development in Night Shift Workers | PDF",1790003661,45]