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Androgen-regulated microRNA-135a decreases prostate cancer cell migration and invasion through downregulating ROCK1 and ROCK2

Oncogene volume 34, pages 2846–2855 (2015)Cite this article

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Abstract

Androgen signaling, via the androgen receptor (AR), is crucial in mediating prostate cancer (PCa) initiation and progression. Identifying new downstream effectors of the androgens/AR pathway will allow a better understanding of these mechanisms and could reveal novel biomarkers and/or therapeutic agents to improve the rate of patient survival. We compared the microRNA expression profiles in androgen-sensitive LNCaP cells stimulated or not with 1 nM R1881 by performing a high-throughput reverse transcriptase–quantitative PCR and found that miR-135a was upregulated. After androgen stimulation, we showed that AR directly activates the transcription of miR-135a2 gene by binding to an androgen response element in the promoter region. Our findings identify miR-135a as a novel effector in androgens/AR signaling. Using xenograft experiments in chick embryos and adult male mice, we showed that miR-135a overexpression decreases in vivo invasion abilities of prostate PC-3 cells. Through in vitro wound-healing migration and invasion assays, we demonstrated that this effect is mediated through downregulating ROCK1 and ROCK2 expression, two genes that we characterized as miR-135a direct target genes. In human surgical samples from prostatectomy, we observed that miR-135a expression was lower in tumoral compared with paired adjacent normal tissues, mainly in tumors classified with a high Gleason score (⩾8). Moreover, miR-135a expression is lower in invasive tumors, showing extraprostatic extension, as compared with intraprostatic localized tumors. In tumor relative to normal glands, we also showed a more frequently higher ROCK1 protein expression determined using a semi-quantitative immunohistochemistry analysis. Therefore, in tumor cells, the lower miR-135a expression could lead to a higher ROCK1 protein expression, which could explain their invasion abilities. The highlighted relationship between miR-135a expression level and the degree of disease aggressiveness suggests that miR-135a may be considered as a prognostic marker in human PCa.

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Acknowledgements

This work was supported by grants from the Institut Mérieux and the Ligue Contre le Cancer. We thank Julien Dugas (CarMen Lab) for his contribution in statistical analyses, Céline Michaux, Karine Castellano and Béatrice Bancel (Service d’Anatomie et Cytologie Pathologiques, Hospices Civils de Lyon) for providing patient samples and performing IHC staining, Jean Viallet for performing in ovo assay, Christophe Chamot and Claire Lionnet (Microscopy core facility) and Patrick Manas (AniRA-PBES platform) of UMS3444/US8 SFR Biosciences Gerland-Lyon Sud, for assistance in wound-healing assay and performing xenograft assay in mice, respectively.

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Authors and Affiliations

  1. Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France

    A Kroiss, S Vincent, M Decaussin-Petrucci, A Ruffion, J Samarut & N Allioli

  2. Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Université Claude Bernard Lyon 1, Pierre-Bénite, France

    M Decaussin-Petrucci, A Ruffion & J Samarut

  3. Service d’Anatomie et de Cytologie Pathologiques, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France

    M Decaussin-Petrucci

  4. Laboratoire CarMen, INSERM U1060, INRA1362, Université Claude Bernard Lyon 1, Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Oullins, France

    E Meugnier

  5. In Ovo, Institut Albert Bonniot, CRI INSERM/UJF U823, Université de Grenoble, La Tronche Cedex, France

    J Viallet

  6. Service d’Urologie, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France

    A Ruffion

  7. GERHM, INSERM U1085-Irset, Université Rennes 1, Rennes, France

    F Chalmel

  8. Service de Biochimie Biologie Moleculaire Sud, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France

    J Samarut

  9. Institut des Sciences Pharmaceutiques et Biologiques (ISPB)-Faculté de Pharmacie de Lyon, Université Claude Bernard Lyon 1, Lyon, France

    N Allioli

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Kroiss, A., Vincent, S., Decaussin-Petrucci, M. et al. Androgen-regulated microRNA-135a decreases prostate cancer cell migration and invasion through downregulating ROCK1 and ROCK2. Oncogene 34, 2846–2855 (2015). https://doi.org/10.1038/onc.2014.222

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