Detection of KRAS gene mutations in endoscopic ultrasound-guided fine-needle aspiration biopsy for improving pancreatic cancer diagnosis

Abstract

Objectives: Endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNAB) is a useful tool in the diagnosis of pancreatic masses. Genetic analysis of these samples could increase the sensitivity and specificity of diagnosis. This study aimed to evaluate the usefulness of a novel method for the detection of mutations in the KRAS (Kirsten rat sarcoma-2 virus) gene for the diagnosis of pancreatic cancer.

Methods: EUS-FNABs were performed on 82 patients with pancreatic masses, including 54 cases of pancreatic ductal adenocarcinoma and 28 of non-malignant pancreatic masses. The biopsies were histopathologically and cytopathologically evaluated, and the detection of KRAS gene mutations (codons 12 and 13) was performed through peptide nucleic acid-directed polymerase chain reaction clamping and DNA sequencing.

Results: In the pancreatic cancer cases, 88.9% (48/54; 95% confidence interval (CI): 80.5-97.2%) had KRAS mutations, while 61.1% (33/54; 95% CI: 48.1-74.1%) were unequivocally diagnosed by histo/cytopathology. In the indeterminate patients (n=49; diagnosed by EUS-FNA as either insufficient material to make a diagnosis, no malignancy, or suspicion of malignancy), there were 10 cases of pancreatic cancer with low serum carbohydrate antigen 19-9 (CA19-9) (<37 U/l) and 6 of these were KRAS mutations. The sensitivity of detection by KRAS mutations (76.2%) and the combination of KRAS mutations and serum CA19-9 (81%) were significantly higher than for serum CA19-9 alone (52.4%). A logistic regression model showed that the KRAS mutation was significant (odds ratio=5.830; CI: 1.531-22.199, P=0.01), but not serum CA19-9. In the non-malignant pancreatic masses (n=28), KRAS mutations were detected in nine precancerous lesions.

Conclusions: Our method for the detection of KRAS gene mutations may be useful to supplement histo/cytopathologic evaluations for pancreatic cancer, and is superior to serum CA19-9 in EUS-FNAB histo/cytopathology-indeterminate patients. Results warrant further verification in other patient populations.