title: A simple method for the routine detection of somatic quantitative genetic alterations in colorectal cancer.
abstract: OBJECTIVE
  Several quantitative genetic alterations have been suggested to have in colorectal cancer (CRC) either a prognostic or a therapeutic predictive value. Routine detection of these alterations is limited by the absence of simple methods.
  
  
  METHODS
  The somatic quantitative multiplex polymerase chain reaction of short fluorescent fragments (QMPSF) is based on the simultaneous amplification under quantitative conditions of several dye-labeled targets both from tumor and nonmalignant tissues. For each patient, the resulting QMPSF fluorescent profiles are superimposed, and quantitative changes are simply detected by an increase or decrease of the corresponding fluorescent peaks. Two assays were developed and applied to 57 CRC: a "bar code" exploring several loci with known prognostic value and a "kinogram" studying the copy number change of kinase genes, against which inhibitors have been developed.
  
  
  RESULTS
  The bar code revealed that the most frequent alterations were the gain of AURKA/20q13 (53%) and MYC/8q24 (39%) and heterozygous deletion of DCC/18q21.3 (39%) and TP53/17p13 (23%). The kinogram detected a gene copy number increase for AURKA, PTK2, MET, and EGFR in 53%, 37%, 33%, and 28% of the tumors, respectively. QMPSF results were validated by comparative genomic hybridization and multiplex real-time polymerase chain reaction on genomic DNA.
  
  
  CONCLUSIONS
  The somatic QMPSF is a simple method able to detect simultaneously on a routine basis several quantitative changes in tumors. Its flexibility will allow the integration of clinically relevant genes. This high throughput method should be a valuable complementary tool of fluorescent in situ hybridization and comparative genomic hybridization.
authors.affiliation: Inserm U614 (IFRMP), University of Rouen and Rouen University Hospital, Institute for Biomedical Research, Rouen, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Killian
authors.forename: Frédéric
authors.initials: F
authors.lastname: Di Fiore
authors.forename: Florence
authors.initials: F
authors.lastname: Le Pessot
authors.forename: France
authors.initials: F
authors.lastname: Blanchard
authors.forename: Aude
authors.initials: A
authors.lastname: Lamy
authors.forename: Grégory
authors.initials: G
authors.lastname: Raux
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Flaman
authors.forename: Bernard
authors.initials: B
authors.lastname: Paillot
authors.forename: Pierre
authors.initials: P
authors.lastname: Michel
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Sabourin
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Tuech
authors.forename: Francis
authors.initials: F
authors.lastname: Michot
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Kerckaert
authors.forename: Richard
authors.initials: R
authors.lastname: Sesboüé
authors.forename: Thierry
authors.initials: T
authors.lastname: Frebourg
mesh_terms: D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D005260:Female; D005453:Fluorescence; D005784:Gene Amplification; D017353:Gene Deletion; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D016163:Genes, DCC / Q000235:genetics; D006801:Humans; D008297:Male; D009367:Neoplasm Staging; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D011379:Prognosis; D011494:Protein Kinases / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D015203:Reproducibility of Results; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D016159:Tumor Suppressor Protein p53; D011494:Protein Kinases