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High stability of microRNAs in tissue samples of compromised quality

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Abstract

Degradation of tissue samples limits performing RNA-based molecular studies, but little is known about the potential usefulness of samples of compromised quality for studies focused on miRNAs. In this work we analyze a series of cryopreserved tissue samples (n = 14), frozen samples that underwent a severe thawing process (n = 10), and their paired formalin-fixed paraffin-embedded (FFPE) tissue samples (n = 24) from patients with breast cancer obtained during primary surgical resection and collected in 2011. Quality and integrity analyses of the total and small fraction of RNA were carried out. Recovery of specific RNA molecules (miRNAs hsa-miR-21, hsa-miR-125b, and hsa-miR-191; snoRNA RNU6B; and mRNAs GAPDH and HPRT1) was also analyzed by quantitative RT-PCR. Our results suggest that visualisation of the small RNA electrophoretic profiles obtained using the Agilent 2100 bioanalyzer makes it possible to differentiate between the three groups of samples (optimally frozen, thawed, and FFPE). We demonstrate that specific miRNA molecules can be similarly recovered from different tissue sample sources, which supports their high degree of stability. We conclude that miRNAs are robustly detected irrespective of the quality of the tissue sample. In this regard, a word of caution should be raised before degraded samples are discarded: although prior quality assessment of the biological material to be analyzed is recommended, our work demonstrates that degraded tissue samples are also suitable for miRNA studies.

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Acknowledgments

This study has been supported by Fundación LeCado-Proyecto Flor de Vida. LPC is funded by the National Health Institute Carlos III FEDER (RD09/0076/00132). MPC is funded by the Generalitat Valenciana Conselleria d'Educació VALi + d program (ACIF/2011/207). GR is funded by the Spanish Ministry of Health (CP08_00069). We want to particularly acknowledge the patients for their participation and INCLIVA Biobank, integrated in the Spanish Hospital Biobanks Network (ReTBioH) and supported by the National Health Institute Carlos III/FEDER and Ministerio de Economía y Competitividad (grant number: RD09/0076/00132). We also thank the Unit for Multigenic Analysis from the Central Unit for Medical Research (UCIM/INCLIVA) for total RNA analysis and the Genomic Unit from Central Service for the Support to Experimental Research (SCSIE) for small RNA analysis.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Biobank, INCLIVA Biomedical Research Institute, Avenida Blasco Ibáñez, 17, 46010, Valencia, Spain

    Lorena Peiró-Chova & Jaime Ferrer-Lozano

  2. Medical Oncology and Hematology, INCLIVA Biomedical Research Institute, Avenida Blasco Ibáñez, 17, Valencia, Spain

    María Peña-Chilet, Elisa Alonso-Yuste, Octavio Burgues, Ana Lluch, Jaime Ferrer-Lozano & Gloria Ribas

  3. Laboratory of Molecular Biology, Fundación Instituto Valenciano de Oncología, C/ Profesor Beltrán Báguena, 8, Valencia, Spain

    José Antonio López-Guerrero

  4. Center for Biomedical Network Research on Rare Diseases, Department of Physiology, School of Medicine and Dentistry, Universitat de Valencia. INCLIVA Biomedical Research Institute, Avenida Blasco Ibáñez, 15, Valencia, Spain

    José Luis García-Giménez

  5. Department of Pathology, Hospital Clínico Universitario de Valencia, Avenida Blasco Ibáñez, 17, Valencia, Spain

    Octavio Burgues

  6. Department of Medical Oncology and Hematology, Hospital Clínico Universitario de Valencia, Avenida Blasco Ibáñez, 17, Valencia, Spain

    Ana Lluch

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  1. Lorena Peiró-Chova
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  2. María Peña-Chilet
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  5. Elisa Alonso-Yuste
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  8. Jaime Ferrer-Lozano
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  9. Gloria Ribas
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Correspondence to Gloria Ribas.

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Peiró-Chova, L., Peña-Chilet, M., López-Guerrero, J.A. et al. High stability of microRNAs in tissue samples of compromised quality. Virchows Arch 463, 765–774 (2013). https://doi.org/10.1007/s00428-013-1485-2

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  • DOI: https://doi.org/10.1007/s00428-013-1485-2

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