Analysis of K‐ras oncogene mutation directly applied to atypical cell clusters on cytologic smear slides of bile and pancreatic juice

To develop an objective reference for the cytological evaluation of atypical cells in bile and pancreatic juice, we analyzed K‐ras oncogene mutation In atypical cell clusters, which were collected directly from cytological smear slides; 50 samples (cell clusters) from 31 smear slides of 21 patients with carcinomas of the pancreatic head region, and nine samples from eight cases of benign disease. These cell clusters (5–1000 cells/cluster) were selectively suspended In buffer containing proteinase K, and subjected to DNA extraction. K‐ras codon 12 mutation was determined by polymerase chain reaction amplification, followed by digestion with BstNI. The K‐ras gene was amplified in 20 of 21 cases with carcinoma (34/50 samples), and In seven of eight cases with non‐neoplastic disease (8/9 samples). Among the cases of which primary tumors showed K‐ras mutation, amplification was successful in 10 of 11 cases; mutation was demonstrated in three of seven cases with cytologically atypical cells (4/11 samples), and in three of three cases with cytologically malignant cells (5/7 samples). No mutation was Identified in the 10 cases of carcinoma without K‐ras mutation (0/15 samples), or in eight cases of non‐neoplastlc disease (0/8 samples). Cytological details could be comparatively evaluated between atypical cell clusters with or without mutation on the same smear slides in two cases. This type of direct analysis of atypical cell clusters may be useful in the self‐assessment of cytological diagnosis of bile and pancreatic juice.

[1]  S. Kini,et al.  Biliary tract cytology in specimens obtained by direct cholangiographic procedures: A study of 74 cases , 1996, Diagnostic cytopathology.

[2]  W. Grody,et al.  Polymerase chain reaction-based K-ras mutation detection of pancreatic adenocarcinoma in routine cytology smears. , 1996, American journal of clinical pathology.

[3]  Scott E. Kern,et al.  DPC4, A Candidate Tumor Suppressor Gene at Human Chromosome 18q21.1 , 1996, Science.

[4]  L. Pradayrol,et al.  Identification of K-ras Mutations in Pancreatic Juice in the Early Diagnosis of Pancreatic Cancer , 1995, Annals of Internal Medicine.

[5]  R. Hruban,et al.  Molecular markers for diagnostic cytology of neoplasms in the head region of the pancreas: mutation of K-ras and overexpression of the p53 protein product. , 1995, Journal of clinical pathology.

[6]  M. Pfreundschuh,et al.  Diagnosis of pancreatic adenocarcinoma by polymerase chain reaction from pancreatic secretions. , 1994, British Journal of Cancer.

[7]  S. Yoshida,et al.  Detection of point mutations in the K‐ras oncogene at codon 12 in pure pancreatic juice for diagnosis of pancreatic carcinoma , 1994, Cancer.

[8]  N. Pellegata,et al.  K-ras and p53 gene mutations in pancreatic cancer: ductal and nonductal tumors progress through different genetic lesions. , 1994, Cancer research.

[9]  S. Maeda,et al.  The role ofras mutation in pancreatic cancer, precancerous lesions, and chronic pancreatitis , 1993 .

[10]  R. Haba,et al.  Detection of c-Ki-ras point mutation from pancreatic juice , 1993, International journal of pancreatology : official journal of the International Association of Pancreatology.

[11]  T. Okai,et al.  Identification of K‐ras Oncogene Mutations in the Pure Pancreatic Juice of Patients with Ductal Pancreatic Cancers , 1993, Japanese journal of cancer research : Gann.

[12]  S. Goodman,et al.  K-ras oncogene activation in adenocarcinoma of the human pancreas. A study of 82 carcinomas using a combination of mutant-enriched polymerase chain reaction analysis and allele-specific oligonucleotide hybridization. , 1993, The American journal of pathology.

[13]  H. Saisho,et al.  Detection of ras gene mutations in pancreatic juice and peripheral blood of patients with pancreatic adenocarcinoma. , 1993, Cancer research.

[14]  S. Hirohashi,et al.  Pancreatic adenocarcinomas frequently show p53 gene mutations. , 1993, The American journal of pathology.

[15]  A. Yanagisawa,et al.  Frequent c-Ki-ras oncogene activation in mucous cell hyperplasias of pancreas suffering from chronic inflammation. , 1993, Cancer research.

[16]  M. Tatsuta,et al.  Cytologic examination of pure pancreatic juice in the diagnosis of pancreatic carcinoma. The endoscopic retrograde intraductal catheter aspiration cytologic technique , 1992, Cancer.

[17]  O. Yokosuka,et al.  Analysis of ras gene mutations in biliary and pancreatic tumors by polymerase chain reaction and direct sequencing , 1990, Cancer.

[18]  D. Shibata,et al.  Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction , 1988, The Journal of experimental medicine.

[19]  K. Mullis,et al.  Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. , 1985, Science.