Effect of radiotherapy on the expression of cardiovascular disease-related miRNA-146a, -155, -221 and -222 in blood of women with breast cancer

RESEARCH ARTICLE Effect of radiotherapy on the expression of cardiovascular disease-related miRNA-146a, -155, -221 and -222 in blood of women with breast cancer Roser Esplugas1,2☯, Meritxell Arenas ID3☯, Noemı́ Serra2, Montserrat Bellés1,2, Marta Bonet ID3, Marina Gascón4, Joan-Carles Vallvé5*, Victoria Linares1,2 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 1 Physiology Unit, School of Medicine, IISPV, Rovira i Virgili University, Reus, Spain, 2 Laboratory of Toxicology and Environmental Health, School of Medicine, IISPV, Rovira i Virgili University, Reus, Spain, 3 Radiation Oncology Department, Sant Joan University Hospital, IISPV, Rovira i Virgili University, Reus, Spain, 4 Radiation Oncology Unit, Miguel Servet University Hospital, Zaragoza, Spain, 5 Research Unit on Lipids and Atherosclerosis, Sant Joan University Hospital, IISPV, Rovira i Virgili University, Reus, Spain ☯ These authors contributed equally to this work. * OPEN ACCESS Citation: Esplugas R, Arenas M, Serra N, Bellés M, Bonet M, Gascón M, et al. (2019) Effect of radiotherapy on the expression of cardiovascular disease-related miRNA-146a, -155, -221 and -222 in blood of women with breast cancer. PLoS ONE 14(5): e0217443. https://doi.org/10.1371/journal. pone.0217443 Editor: Laurent Metzinger, Université de Picardie Jules Verne, FRANCE Received: February 28, 2019 Accepted: May 10, 2019 Published: May 31, 2019 Copyright: © 2019 Esplugas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the manuscript and its Supporting Information files. Funding: This work was supported by grants from the Instituto de Salud Carlos III (PI 10/02547, PI030786), Centro de Investigación Biomédica en Red en Diabetes y Enfermedades Metabólicas asociadas (CIBER-DEM) and Fondo Europeo de Desarrollo Regional (FEDER). The funders had no role in study design, data collection and analysis, Abstract Breast cancer (BC) is one of the most important neoplasias among women. Many patients receive radiotherapy (RT), which involves radiation exposure of the thoracic zone, including the heart and blood vessels, leading to the development of cardiovascular disease (CVD) as a long-term side effect. The severity of CVD-related pathologies leads research on assessing novel CVD biomarkers as diagnostic, prognostic or therapeutic agents. Currently, the possible candidates include blood microRNAs (miRNAs). Previous studies have supported a role for miRNA-146a, -155, -221, and -222 in the progression of CVD. Our purpose was to evaluate the RT-induced modulation of the expression of these miRNAs in the blood of women with BC. Pre-RT control and post-RT blood samples were collected, and after miRNA isolation and reverse transcription, the levels of the selected miRNAs were measured by real-time PCR. Our results showed that miRNA-155 exhibited the lowest expression, while miRNA-222 exhibited the highest expression, followed by miRNA-221. The expression of each individual miRNA was positively correlated with that of the others both pre-RT control and post-RT and inversely correlated with age before RT. Furthermore, RT promoted the overexpression of the selected miRNAs. Their levels were also affected by CVD-linked clinical parameters, treatment and BC side. Modulation of the expression of the selected miRNAs together with other risk factors might be associated with the development of future cardiovascular pathologies. Further confirmatory studies are needed to assess their potential as possible biomarkers in the progression of or as therapeutic targets for RTinduced CVD in BC patients. PLOS ONE | https://doi.org/10.1371/journal.pone.0217443 May 31, 2019 1 / 17 RT modulates CVD-related miRNA-146a, -155, -221 and -222 expression in women with BC decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. Introduction Breast cancer (BC) is one of the most important neoplasias among women. In Spain, 1 in 8 women will develop BC, and approximately 26,000 new cases per year are diagnosed [1]. According to the Instituto Nacional de Epidemiologı́a, there were 6,213 BC deaths in 2014 [2]. Currently, fewer cases in UE and North America are diagnosed because of both early detection and efficient systemic therapies. Nevertheless, it is still the most common cause of death from cancer in less developed countries and the second cause in more developed countries, subsequent to lung cancer [3,4]. The treatment of BC involves a combination of therapies such as surgery, chemotherapy, hormonotherapy, targeted therapy and radiotherapy (RT) [5]. RT can be omitted in some patients with a low-risk profile, which reduces the risk of local recurrence and mortality [6]. RT treatment involves radiation exposure of the thoracic zone, including the heart and blood vessels. Despite RT-related benefits, epidemiological data support a relationship between RT and cardiovascular disease (CVD) [7–9]. Patients with left-sided BC exhibit greater cardiotoxicity than those with right-sided tumours due to RT exposure to the heart and left coronary artery [10–12]. In a retrospective study, Darby et al. [12] observed that for every 1 Gy increase in radiation to the heart, the risk of cardiac damage increased 7.4% in patients with left-sided BC. These authors also reported similar results following ionizing radiation to the heart during RT of BC. Cheng et al. [13], who examined 1,191,371 patients from 39 studies, supported that RT applied to patients with left-sided breast tumours caused major coronary heart failure and cardiac death compared with patients with right-sided BC. They also stated that the risk of CVD begins within the first ten years after RT and that heart mortality increases in the second and third decades. Although the RT-CVD association is lower than that established by risk factors such as smoking, hypertension, diabetes or hyperlipidaemia, the risk of death due to CVD after this therapy exceeds the increase in RT-contributed survival. The CVDs associated with RT in BC patients are coronary heart disease [13,14] and atherosclerosis [15]. The severity of these CVD-related pathologies leads research on assessing novel CVD biomarkers. Currently, the possible candidates as suitable biomarkers in diagnostic, prognostic or therapeutic agents of this pathogenesis are microRNAs (miRNAs) [16–19]. miRNAs are small noncoding RNAs of approximately 22 nucleotides in length that act as posttranscriptional regulators, thus modulating gene expression [20,21]. Approximately 1,500–2,000 human miRNAs have been identified, and it is estimated that over 60% of protein-coding genes are directly regulated by miRNAs. One miRNA targets severa (...truncated)