title: Visualising and quantifying angiogenesis in metastatic colorectal cancer : A comparison of methods and their predictive value for chemotherapy response.
abstract: OBJECTIVE
  Angiogenesis plays an important role in tumour growth and dissemination. We have recently shown that blood vessel density, determined by image analysis based on microRNA-126 (miRNA-126) in situ hybridization (ISH) in the primary tumours of metastatic colorectal cancers (mCRC), is predictive of chemotherapy response. Here, we evaluated whether more general approaches to determine vessel density in primary tumours are equally predictive of chemotherapy response.
  
  
  METHODS
  This methodological study was carried out using paraffin embedded tissues from primary tumours of 89 patients with mCRC, who had all been treated with first-line chemotherapy (XELOX). Tissue sections from the deepest invasive tumour front were processed for miRNA-126 ISH and CD34 immunohistochemistry (IHC). Estimates of microvessel density (MVD) were obtained for both miRNA-126 and CD34 by quantitative image analyses (MVDi), vascular area per image (μm²) analyses, and manually counting vessels in vascular hot spots (MVDh). Clinical responses were evaluated according to Response Evaluation Criteria In Solid Tumours (RECIST).
  
  
  RESULTS
  The MVDi for miRNA-126 showed a significant correlation with treatment response (p=0.01), with a median value of 2,071 μm² (95% CI, 1,505-3,075 μm²) in the responder group compared to 1,337 μm² (95% CI, 1,038-1,499 μm²) in the non-responder group. This difference translated into a significant difference in progression free survival (p=0.01).
  
  
  CONCLUSIONS
  The methodological assessment of MVD and the molecular vessel marker are both important for the prediction of the chemotherapy response in mCRC. Our findings indicate that MVDi for miRNA-126 represents a powerful estimate and may serve as a clinical biomarker superior to MVDh.
authors.affiliation: Department of Oncology, Vejle Hospital, Vejle, Denmark. torben.hansen@rsyd.dk
authors.forename: Torben Frøstrup
authors.initials: TF
authors.lastname: Hansen
authors.forename: Boye Schnack
authors.initials: BS
authors.lastname: Nielsen
authors.forename: Anders
authors.initials: A
authors.lastname: Jakobsen
authors.forename: Flemming Brandt
authors.initials: FB
authors.lastname: Sørensen
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018952:Antigens, CD34 / Q000378:metabolism*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D000069287:Capecitabine; D015179:Colorectal Neoplasms / Q000098:blood supply* / Q000188:drug therapy / Q000473:pathology; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000627:therapeutic use; D018572:Disease-Free Survival; D005260:Female; D005472:Fluorouracil / Q000031:analogs & derivatives / Q000627:therapeutic use; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D055806:Microvessels / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009389:Neovascularization, Pathologic / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D010071:Oxaloacetates; D011379:Prognosis; D055815:Young Adult
chemical_list: D018952:Antigens, CD34; C527111:MIRN126 microRNA, human; D035683:MicroRNAs; D010071:Oxaloacetates; D003841:Deoxycytidine; D000069287:Capecitabine; D005472:Fluorouracil