title: Analysis of Ki-ras gene mutations within the same tumor using a single tumor crypt in colorectal carcinomas.
abstract: BACKGROUND
  The point mutations occurring in codons 12 and 13 of the Ki- ras gene are useful genetic markers to identify intratumoral heterogeneity. A single tumor crypt, which consists of monoclonal cells, can be obtained using the crypt isolation method. Ki- ras gene mutations have been examined using the crypt isolation method to determine whether multiclonarity is present within the same tumor.
  
  
  METHODS
  Ki- ras gene mutations were analyzed using a crypt isolation technique coupled with polymerase chain reaction and direct sequencing in 21 sporadic colorectal carcinomas. The specimens were divided into two groups: a representative sample, which was composed of more than 50 tumor crypts, and a single tumor crypt sample. The latter consisted of 10 single tumor crypts, which were obtained from the same tumor separately.
  
  
  RESULTS
  Ki- ras gene mutations were found in 11 of 21 representative samples and in 12 of 21 single tumor crypt samples. In the 11 samples with Ki- ras mutation, Ki- ras mutations were also found in most single tumor crypts. Among the 12 base substitutions found, G:C to A:T transitions were the most commonly observed. There were no differences between the two samples in the types of Ki- ras mutations found. One Ki- ras mutation that was not detected in the representative sample was observed in only a single tumor crypt.
  
  
  CONCLUSIONS
  Most carcinomas appear to have a homogeneous composition that may result from the successful progression of one of the clones having a Ki- ras mutation. Additional mutations in the Ki- ras gene were rarely observed in colorectal carcinomas.
authors.affiliation: Division of Pathology, Central Clinical Laboratory, Iwate Medical University, 19-1, 020-8505, Morioka, Japan.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Ishii
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Sugai
authors.forename: Wataru
authors.initials: W
authors.lastname: Habano
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Nakamura
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D000328:Adult; D000368:Aged; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D011905:Genes, ras / Q000235:genetics*; D018740:Genetic Heterogeneity; D006801:Humans; D008297:Male; D008875:Middle Aged; D017354:Point Mutation; D017422:Sequence Analysis, DNA
chemical_list: D004273:DNA, Neoplasm