High proportion of mutant K-ras gene in pancreatic juice of patients with pancreatic cystic lesions

Tateishi, Keisuke; Tada, Minoru; Yamagata, Michiko; Isayama, Hiroyuki; Komatsu, Yutaka; Kawabe, Takao; Shiratori, Yasushi; Omata, Masao

doi:10.1136/gut.45.5.737

Cited by 43 publications

(20 citation statements)

References 22 publications

(13 reference statements)

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“…Also, the testing for the presence of mutant KRAS in stool has been proposed for the detection of pancreatic and colorectal carcinoma (32,33). Some studies mentioned other samples like bile or pancreatic juice (34,13,29). Detection of KRAS mutation in pancreatic juice alone is considered insuffi cient for discriminating between pancreatic cancer and benign diseases (35).…”

Section: Discussionmentioning

confidence: 99%

Detection of point mutations in KRAS oncogene by real-time PCR-based genotyping assay in GIT diseases

Ugorcakova

Hlavatý²,

Novotna³

et al. 2012

BLL

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Abstract:Objectives: The determination of gene mutations is important for the diagnosis and prognosis of various gastrointestinal cancers. The aim of our study was to develop a new procedure for the analysis of KRAS gene mutation by application of the real-time PCR method. Background: The detection process requires discriminate trace amount of mutant allele in a large excess of wild-type DNA in various samples. Methods:The real-time PCR based technique using hybridization probes for fi ve most frequently KRAS codon 12 mutations and WT specifi c peptide nucleic acid (PNA) was performed. Our multiplex detection system was tested in various DNA samples (tissue, bile, pancreatic juice) of patients with different diagnoses of gastrointestinal tract disease obtained by endoscopy and ERCP. Results: We designed and optimized the real-time PCR conditions and tested various amount of PNA in PCR reaction to suppress amplifi cation of the wild-type DNA. We determined the interassay variability of the melting temperatures and the results of mutation testing were confi rmed by DNA sequencing with the 100 % accuracy. Incidence of searched mutations was 67.5 % in cohort of 40 patients; for KRAS G12D it was in 44.4 %, KRAS G12V in 22.2 %, KRAS G12S in 14.8 %, KRAS G12A in 14.8 % and KRAS G12C in 3.8 %. The sensitivity of the assays is 1x10 -5 . Conclusions: Advantages of this technique are rapidity, accuracy and it is generally easy to perform. This method can be adapted for synchronic detection of multiple mutations and after readjustment by other type mutation of KRAS gene may serve as useful clinical tool for analyzing point mutations in various clinical samples (Tab. 3, Fig. 3, Ref. 42). Full Text in PDF www.elis.sk.

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Section: Discussionmentioning

confidence: 99%

Detection of point mutations in KRAS oncogene by real-time PCR-based genotyping assay in GIT diseases

Ugorcakova

Hlavatý²,

Novotna³

et al. 2012

BLL

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“…A stepwise increase in K-ras mutation at codon 12 (GGT to GTT or GAT), which is highly associated with pancreatic duct cell carcinoma, has also been found in both IPMT and MCT with increased epithelial dysplasia [2,18,30,33,39,44], although several other studies have shown that K-ras mutation occurs even in a remarkably high percentage of non-neoplastic tissue [3,22,37]. Immunohistochemical overexpression of oncogenes and tumor suppressor genes, such as p53, c-erbB-2, and Dpc-4, has been reported in IPMT [16,19,33].…”

Section: Introductionmentioning

confidence: 96%

Clonality and K- ras mutation analyses of epithelia in intraductal papillary mucinous tumor and mucinous cystic tumor of the pancreas

et al. 2002

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Histological criteria for subclassification of intraductal papillary mucinous tumor (IPMT) and mucinous cystic tumor (MCT) of the pancreas remain ambiguous in the absence of apparent invasion or metastasis. To elucidate this issue, we evaluated clonality and K- ras mutations in 11 cystic tumors of the pancreas from female patients, including 7 IPMTs and 4 MCTs. The analyses were performed on DNA from laser microdissected epithelia showing different degrees of atypia as well as normal-appearing epithelia (NAE) in the individual tumors. The grades of atypia were classified into three groups on conventional hematoxylin-eosin staining. Clonality was assessed using the methylation-induced polymorphic inactivation of the X-linked phosphoglycerate kinase gene. The incidence of monoclonality increased with the grades of atypia: 27% for NAE, 43% for grade 1, and 100% for grades 2 and 3. In three of four MCTs, foci of NAE were polyclonal, while monoclonality was seen in each one of grades 1 and 2. The frequency of K- ras mutation depended on the grades of atypia: 0% for NAE, 29% for grade 1, 50% for grade 2, and 75% for grade 3. Polyclonal epithelia were devoid of K- ras mutation in 92% of sites, while monoclonality was associated with both wild and mutational types in an approximately equal ratio. Both IPMT and MCT seem to arise from polyclonal epithelia and to be replaced by monoclonal neoplastic cells as they undergo dysplastic changes and K- ras mutation. These data suggest that the monoclonal expansion precedes K- ras mutation.

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“…7,10,11 Difficulties in morphological differentiation of CP and PDAC are attributed to extensive fibrosis that is ultimately caused by a variety of genetic alterations, many of which are common between both disease states. [12][13][14][15][16][17] Recent developments using various molecular detection technologies including serial analysis of gene expression, cDNA microarrays, tissue arrays and the human genome-sequencing project has led to the discovery of multiple genes that are overexpressed in CP and PDAC. 4,[17][18][19][20][21][22] In the present study, we selected 29 genes known to be differentially expressed in PDAC or CP from published literature studies and from our own microarray analysis.…”

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confidence: 99%

Accurate discrimination of pancreatic ductal adenocarcinoma and chronic pancreatitis using multimarker expression data and samples obtained by minimally invasive fine needle aspiration

Chen

Zheng

Robbins

et al. 2007

Intl Journal of Cancer

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To augment cytological diagnosis of pancreatic ductal adenocarcinoma (PDAC) in tissue samples obtained by minimally invasive endoscopic ultrasound-guided fine needle aspiration, we investigated whether a small set of molecular markers could accurately distinguish PDAC from chronic pancreatitis (CP). Expression levels of 29 genes were first determined by quantitative real-time RT-PCR in a training set of tissues in which the final diagnosis was PDAC (n 5 20) or CP (n 5 10). Using receiver operator characteristic curve analysis, we determined that the single gene with the highest diagnostic accuracy for discrimination of CP vs. PDAC in the training study was urokinase plasminogen activator receptor (UPAR; AUC value 5 0.895, 95% CI 5 0.728-0.976). In the set of test tissues (n 5 14), the accuracy of UPAR decreased to 79%. However, we observed that the addition of 6 genes (EPCAM2, MAL2, CEA5, CEA6, MSLN and TRIM29; referred to as the 6-gene classifier) to UPAR resulted in high accuracy in both training and testing sets. Excluding 3 samples (out of 44; 7%) for which results of the UPAR/6-gene classifier were ''undefined,'' the accuracy of the UPAR/6-gene classifier was 100% in training samples (n 5 29), 92% in 12 test samples (p 5 0.004 that results were randomly generated; p 5 0.046 that the UPAR/6-gene classifier was comparable to UPAR alone; v 2 test), 100% in 3 samples for which the initial cytological diagnosis was ''suspicious'' and 98% (40/41) overall. Our results provide evidence that molecular marker expression data can be used to augment cytological analysis. ' 2006 Wiley-Liss, Inc.Key words: chronic pancreatitis; pancreatic ductal adenocarcinoma; urokinase plasminogen activator receptor; diagnostic accuracy Although the incidence of pancreatic ductal adenocarcinoma (PDAC) is only 1-2% and relatively low compared to other cancers, nearly all patients die from PDAC within 1-2 years.

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High proportion of mutant K-ras gene in pancreatic juice of patients with pancreatic cystic lesions

Cited by 43 publications

References 22 publications

Detection of point mutations in KRAS oncogene by real-time PCR-based genotyping assay in GIT diseases

Detection of point mutations in KRAS oncogene by real-time PCR-based genotyping assay in GIT diseases

Clonality and K- ras mutation analyses of epithelia in intraductal papillary mucinous tumor and mucinous cystic tumor of the pancreas

Accurate discrimination of pancreatic ductal adenocarcinoma and chronic pancreatitis using multimarker expression data and samples obtained by minimally invasive fine needle aspiration

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