title: Microarray-based identification and RT-PCR test screening for epithelial-specific mRNAs in peripheral blood of patients with colon cancer.
abstract: BACKGROUND
  The efficacy of screening for colorectal cancer using a simple blood-based assay for the detection of tumor cells disseminated in the circulation at an early stage of the disease is gaining positive feedback from several lines of research. This method seems able to reduce colorectal cancer mortality and may replace colonoscopy as the most effective means of detecting colonic lesions.
  
  
  METHODS
  In this work, we present a new microarray-based high-throughput screening method to identifying candidate marker mRNAs for the early detection of epithelial cells diluted in peripheral blood cells. This method includes 1. direct comparison of different samples of colonic mucosa and of blood cells to identify consistent epithelial-specific mRNAs from among 20,000 cDNA assayed by microarray slides; 2. identification of candidate marker mRNAs by data analysis, which allowed selection of only 10 putative differentially expressed genes; 3. Selection of some of the most suitable mRNAs (TMEM69, RANBP3 and PRSS22) that were assayed in blood samples from normal subjects and patients with colon cancer as possible markers for the presence of epithelial cells in the blood, using reverse transcription-polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  Our present results seem to provide an indication, for the first time obtained by genome-scale screening, that a suitable and consistent colon epithelium mRNA marker may be difficult to identify.
  
  
  CONCLUSIONS
  The design of new approaches to identify such markers is warranted.
authors.affiliation: Department of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, I-40126 Bologna, Italy. solmi@alma.unibo.it
authors.forename: Rossella
authors.initials: R
authors.lastname: Solmi
authors.forename: Giampaolo
authors.initials: G
authors.lastname: Ugolini
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Rosati
authors.forename: Simone
authors.initials: S
authors.lastname: Zanotti
authors.forename: Mattia
authors.initials: M
authors.lastname: Lauriola
authors.forename: Isacco
authors.initials: I
authors.lastname: Montroni
authors.forename: Marco
authors.initials: M
authors.lastname: del Governatore
authors.forename: Antonello
authors.initials: A
authors.lastname: Caira
authors.forename: Mario
authors.initials: M
authors.lastname: Taffurelli
authors.forename: Donatella
authors.initials: D
authors.lastname: Santini
authors.forename: Domenico
authors.initials: D
authors.lastname: Coppola
authors.forename: Lia
authors.initials: L
authors.lastname: Guidotti
authors.forename: Paolo
authors.initials: P
authors.lastname: Carinci
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Strippoli
mesh_terms: D000328:Adult; D000368:Aged; D001331:Automation; D003110:Colonic Neoplasms / Q000097:blood*; D004847:Epithelial Cells / Q000737:chemistry*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D053554:Keratin-20; D007633:Keratins / Q000097:blood; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000737:chemistry*; D009687:Nuclear Proteins / Q000235:genetics; D029543:Nucleocytoplasmic Transport Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000097:blood*; D012334:RNA, Neoplasm / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012697:Serine Endopeptidases / Q000235:genetics
chemical_list: C507499:KRT20 protein, human; D053554:Keratin-20; D008565:Membrane Proteins; D009687:Nuclear Proteins; D029543:Nucleocytoplasmic Transport Proteins; C112820:RANBP3 protein, human; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D007633:Keratins; D012697:Serine Endopeptidases; C500893:PRSS22 protein, human