title: The clinical and biological significance of MIR-224 expression in colorectal cancer metastasis.
abstract: MicroRNA (miRNA) expression profile can be used as prognostic marker for human cancers. We aim to explore the significance of miRNAs in colorectal cancer (CRC) metastasis.
  
  
  
  We performed miRNA microarrays using primary CRC tissues from patients with and without metastasis, and validated selected candidates in 85 CRC samples by quantitative real-time PCR (qRT-PCR). We tested metastatic activity of selected miRNAs and identified miRNA targets by prediction algorithms, qRT-PCR, western blot and luciferase assays. Clinical outcomes were analysed in six sets of CRC cases (n=449), including The Cancer Genome Atlas (TCGA) consortium and correlated with miR-224 status. We used the Kaplan-Meier method and log-rank test to assess the difference in survival between patients with low or high levels of miR-224 expression.
  
  
  
  MiR-224 expression increases consistently with tumour burden and microsatellite stable status, and miR-224 enhances CRC metastasis in vitro and in vivo. We identified SMAD4 as a miR-224 target and observed negative correlation (Spearman Rs=-0.44, p<0.0001) between SMAD4 and miR-224 expression in clinical samples. Patients with high miR-224 levels display shorter overall survival in multiple CRC cohorts (p=0.0259, 0.0137, 0.0207, 0.0181, 0.0331 and 0.0037, respectively), and shorter metastasis-free survival (HR 6.51, 95% CI 1.97 to 21.51, p=0.0008). In the TCGA set, combined analysis of miR-224 with SMAD4 expression enhanced correlation with survival (HR 4.12, 95% CI 1.1 to 15.41, p=0.0175).
  
  
  
  MiR-224 promotes CRC metastasis, at least in part, through the regulation of SMAD4. MiR-224 expression in primary CRC, alone or combined with its targets, may have prognostic value for survival of patients with CRC.
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Hui
authors: H
authors: Ling
authors: Cancer Research UK Centre, University of Southampton Cancer Sciences Division, Somers Cancer Research Building, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.
authors: Karen
authors: K
authors: Pickard
authors: Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Cristina
authors: C
authors: Ivan
authors: University of Torino, Department of Oncology, Torino, Italy.|IRCC, Institute for Cancer Research and Treatment, Candiolo, Torino, Italy.
authors: Claudio
authors: C
authors: Isella
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Laboratory of Microbiology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
authors: Mariko
authors: M
authors: Ikuo
authors: Bioinformatics Unit, London Research Institute, Cancer Research UK, London, UK.
authors: Richard
authors: R
authors: Mitter
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Division of Experimental Oncology B, CRO, National Cancer Institute, Aviano, Italy.
authors: Riccardo
authors: R
authors: Spizzo
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Cancer Research UK Centre, University of Southampton Cancer Sciences Division, Somers Cancer Research Building, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.
authors: Marc
authors: M
authors: Bullock
authors: Department of Functional Genomics, The Oncology Institute, Cluj-Napoca, Romania.
authors: Cornelia
authors: C
authors: Braicu
authors: Department of Functional Genomics, The Oncology Institute, Cluj-Napoca, Romania.
authors: Valentina
authors: V
authors: Pileczki
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Kimberly
authors: K
authors: Vincent
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Division of Oncology, Medical University of Graz, Austria.
authors: Martin
authors: M
authors: Pichler
authors: Division of Oncology, Medical University of Graz, Austria.
authors: Verena
authors: V
authors: Stiegelbauer
authors: Institute of Pathology, Medical University of Graz, Austria.
authors: Gerald
authors: G
authors: Hoefler
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|INEB, Instituto de Engenharia Biomedica, University of Porto, Porto, Portugal.
authors: Maria I
authors: MI
authors: Almeida
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Annie
authors: A
authors: Hsiao
authors: Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Xinna
authors: X
authors: Zhang
authors: Cancer Research UK Centre, University of Southampton Cancer Sciences Division, Somers Cancer Research Building, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.|Department of Surgery, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.
authors: John
authors: J
authors: Primrose
authors: Cancer Research UK Centre, University of Southampton Cancer Sciences Division, Somers Cancer Research Building, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.
authors: Graham
authors: G
authors: Packham
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Kevin
authors: K
authors: Liu
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Krishna
authors: K
authors: Bojja
authors: Section of Pathology and Molecular Diagnostics, University of Ferrara, Ferrara, Italy.
authors: Roberta
authors: R
authors: Gafà
authors: Division of Quantitative Science, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA.
authors: Lianchun
authors: L
authors: Xiao
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Simona
authors: S
authors: Rossi
authors: Department of Genitourinary Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Jian H
authors: JH
authors: Song
authors: Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) s.r.l., IRCCS, Gene Therapy Unit, Meldola (FC), Italy.
authors: Ivan
authors: I
authors: Vannini
authors: Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) s.r.l., IRCCS, Gene Therapy Unit, Meldola (FC), Italy.
authors: Francesca
authors: F
authors: Fanini
authors: Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Scott
authors: S
authors: Kopetz
authors: Department of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Patrick
authors: P
authors: Zweidler-McKay
authors: Division of Quantitative Science, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA.
authors: Xuemei
authors: X
authors: Wang
authors: Surgical Clinic 1, Cluj County Hospital, Romania.|UMF Surgery Department 1, Cluj-Napoca, Romania.
authors: Calin
authors: C
authors: Ionescu
authors: Department of Surgical and Gynecology Oncology, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj-Napoca, Romania.
authors: Alexandru
authors: A
authors: Irimie
authors: Departments of Pediatrics, and Molecular Microbiology and Immunology, University of Southern California, Keck School of Medicine, The Saban Research Institute, Children's Center for Cancer and Blood Diseases, Children's Hospital Los Angeles, Los Angeles, CA, USA.|Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) s.r.l., IRCCS, Gene Therapy Unit, Meldola (FC), Italy.
authors: Muller
authors: M
authors: Fabbri
authors: Section of Pathology and Molecular Diagnostics, University of Ferrara, Ferrara, Italy.
authors: Giovanni
authors: G
authors: Lanza
authors: Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: Stanley R
authors: SR
authors: Hamilton
authors: Department of Functional Genomics, The Oncology Institute, Cluj-Napoca, Romania.|Research Center for Functional Genomics, Biomedicine and Translational Medicine, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj-Napoca, Romania.
authors: Ioana
authors: I
authors: Berindan-Neagoe
authors: University of Torino, Department of Oncology, Torino, Italy.|IRCC, Institute for Cancer Research and Treatment, Candiolo, Torino, Italy.
authors: Enzo
authors: E
authors: Medico
authors: Cancer Research UK Centre, University of Southampton Cancer Sciences Division, Somers Cancer Research Building, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.|Department of Surgery, Southampton University Hospital NHS Trust, Tremona road, Southampton, SO16 6YD, UK.
authors: Alex
authors: A
authors: Mirnezami
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors: George A
authors: GA
authors: Calin
authors: Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.|Division of Experimental Oncology B, CRO, National Cancer Institute, Aviano, Italy.
authors: Milena S
authors: MS
authors: Nicoloso
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000235:genetics / Q000401:mortality / Q000556:secondary; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001317:Austria; D014408:Biomarkers, Tumor / Q000097:blood*; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D066298:In Vitro Techniques; D007558:Italy; D053208:Kaplan-Meier Estimate; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000097:blood*; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D011237:Predictive Value of Tests; D012383:Romania; D012680:Sensitivity and Specificity; D006113:United Kingdom
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs
keywords: COLORECTAL CANCER; LIVER METASTASES; MICROSATELLITE INSTABILITY; MOLECULAR GENETICS; RNA EXPRESSION