Department of Gastroenterological Surgery (Surgery II), Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi, 466-8560, Japan.
Clin Epigenetics. 2021 Sep 16;13(1):172. doi: 10.1186/s13148-021-01165-8.
Pancreatic cancer is one of the lethal cancers among solid malignancies. Pathological diagnosis of surgical margins is sometimes unreliable due to tissue shrinkage, invisible field cancerization and skipped lesions like tumor budding. As a result, tumor recurrences sometimes occur even from the pathologically negative surgical margins.
We applied molecular surgical margin (MSM) analysis by tissue imprinting procedure to improve the detection sensitivity of tiny cancerous cells on the surgical specimen surface after pancreatoduodenectomy. Surgical specimens were collected from 45 pancreatic cancer cases who received subtotal stomach preserving pancreatoduodenectomy at Nagoya University Hospital during 2017-2019. Quantitative methylation-specific PCR (QMSP) of the original methylation marker panel (CD1D, KCNK12, PAX5) were performed and analyzed with postoperative survival outcomes.
Among 45 tumors, 26 cases (58%) were QMSP-positive for CD1D, 25 (56%) for KCNK12 and 27 (60%) for PAX5. Among the 38 tumors in which at least one of the three markers was positive, CD1D-positive cancer cells, KCNK12-positive cancer cells, and PAX5-positive cancer cells were detected at the surgical margin in 8 cases, 7 cases and 10 cases, respectively. Consequently, a total of 17 patients had at least one marker detected at the surgical margin by QMSP, and these patients were defined as MSM-positive. They were associated with significantly poor recurrence-free survival (p = 0.002) and overall survival (p = 0.005) than MSM-negative patients. Multivariable analysis showed that MSM-positive was the only significant independent factor for worse recurrence-free survival (hazard ratio: 3.522, 95% confidence interval: 1.352-9.179, p = 0.010). On the other hand, a significant proportion of MSM-negative cases were found to have received neoadjuvant chemotherapy (p = 0.019).
Pancreatic cancer-specific methylation marker panel was established to perform MSM analysis. MSM-positive status might represent microscopically undetectable cancer cells on the surgical margin and might influence the postoperative long-term outcomes.
胰腺癌是实体恶性肿瘤中致命的癌症之一。由于组织收缩、不可见的场癌变和肿瘤芽等 skipped lesions，手术切缘的病理诊断有时不可靠。因此，即使手术切缘病理阴性，肿瘤也会复发。
我们应用组织印迹法进行分子外科切缘（MSM）分析，以提高胰十二指肠切除术后手术标本表面微小癌细胞的检测灵敏度。收集了 2017 年至 2019 年期间在名古屋大学医院接受部分胃保留胰十二指肠切除术的 45 例胰腺癌患者的手术标本。对原始甲基化标志物（CD1D、KCNK12、PAX5）进行定量甲基化特异性 PCR（QMSP）分析，并结合术后生存结果进行分析。
在 45 例肿瘤中，26 例（58%）CD1D、25 例（56%）KCNK12 和 27 例（60%）PAX5 的 QMSP 阳性。在至少有一个标志物阳性的 38 例肿瘤中，8 例、7 例和 10 例在手术切缘处检测到 CD1D 阳性癌细胞、KCNK12 阳性癌细胞和 PAX5 阳性癌细胞。因此，共有 17 例患者的 QMSP 在手术切缘处检测到至少一个标志物，这些患者被定义为 MSM 阳性。与 MSM 阴性患者相比，他们的无复发生存率（p=0.002）和总生存率（p=0.005）显著更差。多变量分析显示，MSM 阳性是无复发生存率更差的唯一显著独立因素（危险比：3.522，95%置信区间：1.352-9.179，p=0.010）。另一方面，发现相当比例的 MSM 阴性病例接受了新辅助化疗（p=0.019）。
建立了胰腺癌特异性甲基化标志物panel 进行 MSM 分析。MSM 阳性状态可能代表手术切缘处无法检测到的显微镜下癌细胞，并可能影响术后长期结局。
