title: Androgen-regulated microRNA-135a decreases prostate cancer cell migration and invasion through downregulating ROCK1 and ROCK2.
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.
authors: Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France.
authors: A
authors: A
authors: Kroiss
authors: Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France.
authors: S
authors: S
authors: Vincent
authors: 1] Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France [2] Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Université Claude Bernard Lyon 1, Pierre-Bénite, France [3] Service d'Anatomie et de Cytologie Pathologiques, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France.
authors: M
authors: M
authors: Decaussin-Petrucci
authors: Laboratoire CarMen, INSERM U1060, INRA1362, Université Claude Bernard Lyon 1, Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Oullins, France.
authors: E
authors: E
authors: Meugnier
authors: In Ovo, Institut Albert Bonniot, CRI INSERM/UJF U823, Université de Grenoble, La Tronche Cedex, France.
authors: J
authors: J
authors: Viallet
authors: 1] Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France [2] Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Université Claude Bernard Lyon 1, Pierre-Bénite, France [3] Service d'Urologie, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France.
authors: A
authors: A
authors: Ruffion
authors: GERHM, INSERM U1085-Irset, Université Rennes 1, Rennes, France.
authors: F
authors: F
authors: Chalmel
authors: 1] Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France [2] Faculté de Médecine et de Maïeutique Lyon-Sud-Charles Mérieux, Université Claude Bernard Lyon 1, Pierre-Bénite, France [3] Service de Biochimie Biologie Moleculaire Sud, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, Pierre-Bénite, France.
authors: J
authors: J
authors: Samarut
authors: 1] Université de Lyon, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Institut de Génomique Fonctionnelle de Lyon (IGFL), Lyon, France [2] Institut des Sciences Pharmaceutiques et Biologiques (ISPB)-Faculté de Pharmacie de Lyon, Université Claude Bernard Lyon 1, Lyon, France.
authors: N
authors: N
authors: Allioli
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology*; D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics*; D002642:Chick Embryo; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology*; D054460:rho-Associated Kinases / Q000235:genetics*
chemical_list: D000728:Androgens; C544965:MIRN135 microRNA, human; D035683:MicroRNAs; C515851:ROCK1 protein, human; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases