Theranostics 2018; 8(8):2147-2160. doi:10.7150/thno.22726

Research Paper

Release of MicroRNAs into Body Fluids from Ten Organs of Mice Exposed to Cigarette Smoke

Alberto Izzotti1,2, Mariagrazia Longobardi1, Sebastiano La Maestra1, Rosanna T. Micale1, Alessandra Pulliero1, Anna Camoirano1, Marta Geretto1, Francesco D'Agostini1, Roumen Balansky1,3, Mark Steven Miller4, Vernon E. Steele4, Silvio De Flora1✉

1. Department of Health Sciences, University of Genoa, 16132 Genoa, Italy
2. IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy
3. National Center of Oncology, Sofia-1756, Bulgaria
4. Division of Cancer Prevention, National Cancer Institute, Rockville, MD 20850, USA

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license ( See for full terms and conditions.
Izzotti A, Longobardi M, La Maestra S, Micale RT, Pulliero A, Camoirano A, Geretto M, D'Agostini F, Balansky R, Miller MS, Steele VE, De Flora S. Release of MicroRNAs into Body Fluids from Ten Organs of Mice Exposed to Cigarette Smoke. Theranostics 2018; 8(8):2147-2160. doi:10.7150/thno.22726. Available from

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Purpose: MicroRNAs are small non-coding RNAs that regulate gene expression, thereby playing a role in a variety of physiological and pathophysiological states. Exposure to cigarette smoke extensively downregulates microRNA expression in pulmonary cells of mice, rats, and humans. Cellular microRNAs are released into body fluids, but a poor parallelism was previously observed between lung microRNAs and circulating microRNAs. The purpose of the present study was to validate the application of this epigenetic biomarker by using less invasive collection procedures.

Experimental design: Using microarray analyses, we measured 1135 microRNAs in 10 organs and 3 body fluids of mice that were either unexposed or exposed to mainstream cigarette smoke for up to 8 weeks. The results obtained with selected miRNAs were validated by qPCR.

Results: The lung was the main target affected by smoke (190 dysregulated miRNAs), followed by skeletal muscle (180), liver (138), blood serum (109), kidney (96), spleen (89), stomach (36), heart (33), bronchoalveolar lavage fluid (32), urine (27), urinary bladder (12), colon (5), and brain (0). Skeletal muscle, kidney, and lung were the most important sources of smoke-altered microRNAs in blood serum, urine, and bronchoalveolar lavage fluid, respectively.

Conclusions: microRNA expression analysis was able to identify target organs after just 8 weeks of exposure to smoke, well before the occurrence of any detectable histopathological alteration. The present translational study validates the use of body fluid microRNAs as biomarkers applicable to human biomonitoring for mechanistic studies, diagnostic purposes, preventive medicine, and therapeutic strategies.

Keywords: microRNAs, cigarette smoke, interorgan distribution, body fluids