K-ras mutations in duodenal aspirate without secretin stimulation for screening of pancreatic and biliary tract carcinoma

Watanabe, Hiroyuki; Ha, Aiguli; Hu, Yuxin; Ohtsubo, Koushiro; Yamaguchi, Yasushi; Motoo, Yoshiharu; Okai, Takashi; Toya, Daishu; Tanaka, Nobuyoshi; Sawabu, Norio

doi:10.1002/(sici)1097-0142(19991015)86:8<1441::aid-cncr9>3.0.co;2-i

Cited by 40 publications

(27 citation statements)

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“…Amplifications were performed in 50 L of reaction mixture under the following conditions: 1 cycle at 95 ℃ for 360 s, 35 cycles at 94 ℃ for 45 s, 59 ℃ 45 s, and 72 ℃ for 60 s; and a final extension at 72 ℃ for 600 s. Amplified products were visualized by 8% polyacrylamide gel electrophoresis. Mutant allele-specific amplification (MASA) G to A mutations at codon 12 (GGT) were detected by MASA [12] . The 3'-terminal bases of sense primers were set to the mutant base of K-ras codon 12; the sense-1 primer was for the first base G to A mutation and the sense-2 primer was for the second.…”

Section: Methylation-specific Pcr (Msp)mentioning

confidence: 99%

Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Zhu²,

Huang³

et al. 2005

WJG

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AIM:To investigate the functions of promoter hypermethylation of O 6 -methylguanine-DNA methyltransferase (MGMT) gene in colorectal tumorigenesis and progression. METHODS:The promoter hypermethylation of MGMT gene was detected in 27 sporadic colorectal adenomas, 62 sporadic colorectal carcinomas and 20 normal colorectal mucosa tissues by methylation-specific PCR. At the same time, the expression of MGMT protein was carried out in the same samples using immunohistochemistry. Mutantallele-specific amplification was used to detect K-ras G to A point mutation in codon 12. RESULTS:None of the normal colorectal mucosa tissues showed methylated bands. Promoter hypermethylation was detected in 40.7% (11 of 27) of adenomas and 43.5% (27 of 62) of carcinomas. MGMT proteins were expressed in nucleus and cytoplasm of normal colorectal mucosa tissues. Loss of MGMT expression was found in 22.2% (6 of 27) of adenomas and 45.2% (28 of 62) of carcinomas. The difference between them was significant (P = 0.041). In the 6 adenomas and 28 carcinomas losing MGMT expression, 5 and 24 cases presented methylation, respectively (P = 0.027, P<0.001). Thirteen of the 19 colorectal tumors with K-ras G to A point mutation in codon 12 had methylated MGMT (P = 0.011). The frequencies of K-ras G to A point mutation were 35.3% (12 of 34) and 12.7% (7 of 55) in tumors losing MGMT expression and with normal expression, respectively. CONCLUSION:

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Section: Methylation-specific Pcr (Msp)mentioning

confidence: 99%

Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Zhu²,

Huang³

et al. 2005

WJG

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“…The sensitivity (47%) was in agreement with previous studies (27 to 81%) (Mulcahy et al, 1998;Yamada et al, 1998;Castells et al, 1999;Porta et al, 1999;Theodor et al, 2000;Zambon et al, 2000), although the search for five other possible KRAS2 mutations in codon 12 was not performed. Higher KRAS2 mutation prevalences have been reported in pancreatic or duodenal juice (63 to 87%), due probably to higher DNA tumour content in pancreatic juice as compared to plasma (Wilentz et al, 1998;Van Laethem et al, 1998;Watanabe et al, 1999). Use of samples of pancreatic juice however requires invasive procedures (fine-needle aspiration during endoscopic ultrasonography or endoscopic retrograde pancreatography).…”

Section: Molecular and Cellular Pathologymentioning

confidence: 99%

“…Detection of KRAS2 mutations were first reported in surgically removed pancreatic tumoural tissue or at autopsy (Almoguera et al, 1988;Tada et al, 1991). Thereafter mutations were discovered in 63 to 83% of samples of pure pancreatic juice or main pancreatic duct brushing obtained during endoscopic retrograde pancreatography (Iguchi et al, 1996;Kondo et al, 1997;Tada et al, 1998;Van Laethem et al, 1998;Okai et al, 1999;Watanabe et al, 1999;Ha et al, 2001;Pugliese et al, 2001;Seki et al, 2001) or at fine-needle tumour aspiration (Pabst et al, 1999;Puig et al, 2000), and in 20 to 54% of stools (Caldas et al, 1994;Wenger et al, 1999) from patients with pancreatic cancer. Circulating deoxyribo nucleic acid (DNA) was first detected in serum or plasma of normal subjects in 1975 (Steinman, 1975).…”

mentioning

confidence: 99%

Differential diagnosis between chronic pancreatitis and pancreatic cancer: value of the detection of KRAS2 mutations in circulating DNA

et al. 2002

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KRAS2 mutations in codon 12 have been detected in about 80% of pancreatic cancers. The aim of this study was to evaluate the value of KRAS2 mutations detection in circulating deoxyribo nucleic acid to differentiate pancreatic cancer from chronic pancreatitis. Circulating deoxyribo nucleic acid was isolated from serum in 47 patients with histologically proven pancreatic adenocarcinomas (26 males, median age 65 years) and 31 controls with chronic pancreatitis (26 males, median age 48 years). Mutations at codon 12 of KRAS2 gene were searched for using polymerase chain reaction and allele specific amplification. Serum carbohydrate antigen 19.9 levels were also determined. KRAS2 mutations were found in 22 patients (47%) with pancreatic cancer and in four controls with chronic pancreatitis (13%) (P50.002). None of the latter developed a pancreatic cancer within the 36 months of median follow-up. The sensitivity, specificity, positive and negative predictive values of serum serum KRAS2 mutations for the diagnosis of pancreatic cancer were 47, 87, 85 and 52%, respectively. KRAS2 mutations were not related to age, gender, smoking habit, tumour stage, or survival. Among the 26 patients with normal or non-contributive (due to cholestasis) serum carbohydrate antigen 19.9 levels, 14 (54%) had KRAS2 mutations. The combination of KRAS2 and carbohydrate antigen 19.9 gave a sensitivity, specificity, positive and negative predictive values for the diagnosis of pancreatic cancer of 98, 77, 87 and 96%, respectively. Detection of KRAS2 mutations in circulating deoxyribo nucleic acid has a low sensitivity but a specificity about 90% for the diagnosis of pancreatic cancer. It seems particularly useful when serum carbohydrate antigen 19.9 levels are normal or inconclusive. A combined normal serum carbohydrate antigen 19.9 and absence of circulating KRAS2 mutations makes the diagnosis of pancreatic cancer extremely unlikely.

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“…A K-ras oncogene mutation at codon 12 (KRM) in the cystic fluid was not detected by mutant allele-specific amplification, performed according to our previous report. 13 Superselective gastroduodenal arteriography showed an avascular area concordant with the cystic lesion, without tumor staining or arterial encasement (data not shown). Thus, we made a preoperative diagnosis of macrocystic SCA of the pancreas causing obstructive jaundice, based on the well-demarcated oligocysts, which were 3-4 cm in diameter with a stellate septum and serous contents.…”

Section: Introductionmentioning

confidence: 85%

Successful Cystic Fenestration for a Macrocystic Serous Cystadenoma of the Pancreas Causing Obstructive Jaundice: Report of a Case

et al. 2005

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A 72-year-old man was admitted to our hospital for investigation of jaundice. We made a preoperative diagnosis of macrocystic serous cystadenoma (SCA) of the pancreas, but did not perform palliative choledochojejunostomy because aspiration of the pancreatic cystic fluid caused the cysts to shrink and relieved the compressive stenosis of the common bile duct (CBD) during the operation. Frozen sections of the cyst wall taken by incision biopsy showed no signs of malignancy. Therefore, we performed fenestration of the cystic wall after fixing the inner epithelium of the cyst with 100% ethanol and aspirating the cystic fluid. Cholangiography after the cystic fenestration showed resolution of the CBD stenosis and abdominal computed tomography (CT) confirmed the disappearance of the pancreatic cysts. No recurrence of cystic swelling or obstructive jaundice has been detected by abdominal CT or laboratory data for more than 2 years since the cystic fenestration. Thus, cystic fenestration may be a better palliative option for treating benign compressive tumors such as macrocystic SCA of the pancreas causing obstructive jaundice.

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K-ras mutations in duodenal aspirate without secretin stimulation for screening of pancreatic and biliary tract carcinoma

Cited by 40 publications

References 35 publications

Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Differential diagnosis between chronic pancreatitis and pancreatic cancer: value of the detection of KRAS2 mutations in circulating DNA

Successful Cystic Fenestration for a Macrocystic Serous Cystadenoma of the Pancreas Causing Obstructive Jaundice: Report of a Case

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K-ras mutations in duodenal aspirate without secretin stimulation for screening of pancreatic and biliary tract carcinoma

Cited by 40 publications

References 35 publications

Hypermethylation of CpG island in O6-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Hypermethylation of CpG island in O6-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Differential diagnosis between chronic pancreatitis and pancreatic cancer: value of the detection of KRAS2 mutations in circulating DNA

Successful Cystic Fenestration for a Macrocystic Serous Cystadenoma of the Pancreas Causing Obstructive Jaundice: Report of a Case

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Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor

Hypermethylation of CpG island in O⁶-methylguanine-DNA methyltransferase gene was associated withK-rasG to A mutation in colorectal tumor