Comparison of Native Aspirates and Cytological Smears Obtained by EUS-Guided Biopsies for Effective DNA/RNA Marker Testing in Pancreatic Cancer

Pathol Oncol Res. 2020 Jan;26(1):379-385. doi: 10.1007/s12253-018-0490-9. Epub 2018 Oct 25.

Abstract

We compare two types of pancreatic carcinoma samples obtained by EUS-guided fine needle biopsy (EUS-FNB) in terms of the success rates and clinical validity of analysis of two most commonly investigated DNA/RNA pancreatic cancer markers, KRAS mutations and miR-21 expression. 118 patients with pancreatic ductal adenocarcinoma underwent EUS-FNB. The collected sample was divided, one part was stored in a stabilizing solution as native aspirate (EUS-FNA) and second part was processed into the cytological smear (EUS-FNC). DNA/RNA extraction was followed by analysis of KRAS mutations and miR-21 expression. For both sample types, the yields of DNA/RNA extraction and success rates of KRAS mutation and miRNA expression were evaluated. Finally, the resulting KRAS mutation frequency and miR-21 prognostic role were compared to literature data from tissue resections. The overall amount of isolated DNA/RNA from EUS-FNC was lower compared to the EUS-FNA, average yield 10 ng vs 147 ng for DNA and average yield 164 vs. 642 ng for RNA, but the success rates for KRAS and miR-21 analysis was 100% for both sample types. The KRAS-mutant detection frequency in EUS-FNC was 12% higher than in EUS-FNA (90 vs 78%). The prognostic role of miR-21 was confirmed in EUS-FNC (p = 0.02), but did not reach statistical significance in EUS-FNA (p = 0.06). Although both types of EUS-FNB samples are suitable for DNA/RNA extraction and subsequent DNA mutation and miRNA expression analysis, reliable results with clinical validity were only obtained for EUS-FNC.

Keywords: EUS-FNA; KRAS; Pancreatic cancer; miR-21.

Publication types

  • Comparative Study

MeSH terms

  • Aged
  • Biomarkers, Tumor / analysis*
  • Carcinoma, Pancreatic Ductal / diagnosis*
  • Cytodiagnosis / methods*
  • DNA / analysis
  • Endoscopic Ultrasound-Guided Fine Needle Aspiration
  • Female
  • Humans
  • Male
  • MicroRNAs / analysis
  • Middle Aged
  • Mutation
  • Pancreatic Neoplasms / diagnosis*
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Specimen Handling / methods*
  • Tissue Fixation / methods

Substances

  • Biomarkers, Tumor
  • KRAS protein, human
  • MIRN21 microRNA, human
  • MicroRNAs
  • DNA
  • Proto-Oncogene Proteins p21(ras)