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Specific alterations of the microRNA transcriptome and global network structure in colorectal cancer after treatment with MAPK/ERK inhibitors

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Abstract

The mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway has a master control role in various cancer-related biological processes as cell growth, proliferation, differentiation, migration, and apoptosis. It also regulates many transcription factors that control microRNAs (miRNAs) and their biosynthetic machinery. To investigate on the still poorly characterised global involvement of miRNAs within the pathway, we profiled the expression of 745 miRNAs in three colorectal cancer (CRC) cell lines after blocking the pathway with three different inhibitors. This allowed the identification of two classes of post-treatment differentially expressed (DE) miRNAs: (1) common DE miRNAs in all CRC lines after treatment with a specific inhibitor (class A); (2) DE miRNAs in a single CRC line after treatment with all three inhibitors (class B). By determining the molecular targets, biological roles, network position of chosen miRNAs from class A (miR-372, miR-663b, miR-1226*) and class B (miR-92a-1*, miR-135b*, miR-720), we experimentally demonstrated that they are involved in cell proliferation, migration, apoptosis, and globally affect the regulation circuits centred on MAPK/ERK signaling. Interestingly, the levels of miR-92a-1*, miR-135b*, miR-372, miR-720 are significantly higher in biopsies from CRC patients than in normal controls; they also are significantly higher in CRC patients with mutated KRAS than in those with wild-type genotypes (Wilcoxon test, p < 0.05): the latter could be a downstream effect of ERK pathway overactivation, triggered by KRAS mutations. Finally, our functional data strongly suggest the following miRNA/target pairs: miR-92a-1*/PTEN-SOCS5; miR-135b*/LATS2; miR-372/TXNIP; miR-663b/CCND2. Altogether, these results contribute to deepen current knowledge on still uncharacterized features of MAPK/ERK pathway, pinpointing new oncomiRs in CRC and allowing their translation into clinical practice and CRC therapy.

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Acknowledgments

Prof Michele Purrello and the Researchers of the Unità di BioMedicina Molecolare Genomica e dei Sistemi Complessi, Genetica, Biologia Computazionale are grateful to Prof Karl Grzeschik (Marburg University, Marburg, Germany, UE), Prof Bud Mishra (New York University, New York, NY, USA), Prof Riccardo Vigneri (Università di Catania, Catania, Italy, EU) for their interest toward their studies along the years. We acknowledge the collaboration of Miss R Passanisi and Miss R Pellegrino (students from Corso LT di Scienze Biologiche, Università di Catania) at the time of their thesis preparation. This project was supported by funds from Ministero dell’Università e della Ricerca Scientifica e Tecnologica to Prof M Purrello (FAR 2007: Generation of a technological platform to study the effects of antineoplastic drugs and to investigate their potential efficacy as neuroprotective agents; PRA 2007: Caratterizzazione delle Omiche del Macchinario Apoptotico e dell’Apparato di Trascrizione: ruolo biologico dei microRNA e loro coinvolgimento in Patologia; Progetti Dipartimentali della Facoltà di Medicina e Chirurgia: BioMedicina Molecolare dei Sistemi Complessi: Analisi Molecolare delle Omiche nel Carcinoma del Colon e del Retto ed Applicazioni Cliniche), and PRA to Prof F Basile, A Cappellani, M Di Vita, S Lanzafame.

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Authors and Affiliations

  1. Dipartimento Gian Filippo Ingrassia, Unità di BioMedicina Molecolare Genomica e dei Sistemi Complessi, Genetica, Biologia Computazionale, Università di Catania, Via Santa Sofia 87, 95123, Catania, Italy

    Marco Ragusa, Luisa Statello, Marco Maugeri, Alessandra Majorana, Davide Barbagallo, Loredana Salito, Mariangela Sammito, Manuela Santonocito, Rosario Angelica, Marina Scalia, Cinzia Di Pietro & Michele Purrello

  2. Dipartimento di Chirurgia, Università di Catania, Catania, 95123, Italy

    Andrea Cavallaro, Antonio Biondi, Maria Di Vita, Alessandro Cappellani & Francesco Basile

  3. Dipartimento Gian Filippo Ingrassia, Unità di Anatomia Patologica, Università di Catania, Catania, 95123, Italy

    Rosario Caltabiano, Enrico Vasquez & Salvatore Lanzafame

  4. Dipartimento di Ostetricia, Ginecologia e Scienze Radiologiche, Università di Catania, Catania, 95123, Italy

    Giuseppe Privitera

  5. European Drug Safety and Metabolism Research Center, Myrmex, Catania, 95030, Italy

    Elisabetta Tendi & Salvatore Celeste

Authors
  1. Marco Ragusa
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  2. Luisa Statello
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  3. Marco Maugeri
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  4. Alessandra Majorana
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  5. Davide Barbagallo
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  6. Loredana Salito
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  7. Mariangela Sammito
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  9. Rosario Angelica
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  10. Andrea Cavallaro
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  11. Marina Scalia
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  12. Rosario Caltabiano
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  13. Giuseppe Privitera
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  14. Antonio Biondi
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  15. Maria Di Vita
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  16. Alessandro Cappellani
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  17. Enrico Vasquez
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  18. Salvatore Lanzafame
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  19. Elisabetta Tendi
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  20. Salvatore Celeste
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  21. Cinzia Di Pietro
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  22. Francesco Basile
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  23. Michele Purrello
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Corresponding authors

Correspondence to Marco Ragusa or Michele Purrello.

Additional information

Cinzia Di Pietro, Francesco Basile and Michele Purrello are senior authors.

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Ragusa, M., Statello, L., Maugeri, M. et al. Specific alterations of the microRNA transcriptome and global network structure in colorectal cancer after treatment with MAPK/ERK inhibitors. J Mol Med 90, 1421–1438 (2012). https://doi.org/10.1007/s00109-012-0918-8

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  • DOI: https://doi.org/10.1007/s00109-012-0918-8

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