Centre for Applied Genomics of Solid Tumors (CEGES), Genomac Research Institute, 161 00, Prague, CZ, Czech Republic.
Department of Internal Medicine, 1st Faculty of Medicine of Charles University and Military University Hospital, 169 02, Prague, CZ, Czech Republic.
Pathol Oncol Res. 2020 Jan;26(1):379-385. doi: 10.1007/s12253-018-0490-9. Epub 2018 Oct 25.
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.
我们比较了两种通过超声内镜引导下细针活检(EUS-FNB)获得的胰腺癌样本,以评估两种最常研究的胰腺癌细胞标记物 KRAS 突变和 miR-21 表达的分析成功率和临床有效性。118 例胰腺导管腺癌患者接受了 EUS-FNB。收集的样本分为两部分,一部分储存在稳定溶液中作为原生抽吸物(EUS-FNA),另一部分加工成细胞学涂片(EUS-FNC)。然后进行 DNA/RNA 提取,分析 KRAS 突变和 miR-21 表达。对于两种样本类型,均评估了 DNA/RNA 提取的产量和 KRAS 突变和 miRNA 表达的成功率。最后,将所得 KRAS 突变频率和 miR-21 预后作用与组织切除的文献数据进行比较。与 EUS-FNA 相比,EUS-FNC 中分离的 DNA/RNA 总量较低,DNA 的平均产量为 10 ng 对 147 ng,RNA 的平均产量为 164 ng 对 642 ng,但两种样本类型的 KRAS 和 miR-21 分析成功率均为 100%。EUS-FNC 中 KRAS 突变检测频率比 EUS-FNA 高 12%(90%比 78%)。miR-21 的预后作用在 EUS-FNC 中得到证实(p=0.02),但在 EUS-FNA 中未达到统计学意义(p=0.06)。尽管两种类型的 EUS-FNB 样本均适用于 DNA/RNA 提取以及随后的 DNA 突变和 miRNA 表达分析,但仅在 EUS-FNC 中获得了具有临床有效性的可靠结果。