Molecular Detection of Sentinel Node Micrometastases in Patients with Clinical N0 Gastric Carcinoma with Real-time Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Abstract

The real-time multiplex RT-PCR assay is a useful tool for the detection of MM in SNs and NSNs in patients with gastric cancer.

A new stage of sentinel node navigation surgery in early gastric cancer

“…It was reported that 24 % of patients with SN metastasis diagnosed by permanent sections could not be identified by frozen sections during the operation in the trial conducted by the Japan Society of Sentinel Node Navigation Surgery [7]. Different methods such as multistep level section, immunohistochemistry [23,24], reverse transcription PCR (RT-PCR) [25][26][27][28], and the one-step nucleic acid amplification assay [29] have been developed to reduce the falsenegative rate by an additional diagnosis for micrometastasis in SNs. Micrometastases were detected in four of 35 patients (11 %), and six of 1,028 nodes (0.58 %) without metastasis diagnosed by permanent section, using a combined technique of complete serial sectioning and immunohistochemistry [23].…”

“…Micrometastases were detected in four of 35 patients (11 %), and six of 1,028 nodes (0.58 %) without metastasis diagnosed by permanent section, using a combined technique of complete serial sectioning and immunohistochemistry [23]. Shimizu et al [28] established a highly sensitive real-time RT-PCR system to detect the messenger RNA (mRNA) of cytokeratin 19, cytokeratin 20, and carcinoembryonic antigen, demonstrating that 27 % of the patients had negative pathological but positive RT-PCR findings. In seven patients (6.8 %), SNs were negative but non-SNs were positive on RT-PCR in that report.…”

“…In this sense, the falsenegative rates might not reach 0 % even if there are advancements in molecular biological methods in the future. But almost articles about micrometastasis in SN biopsy support the oncological safety of LBD [23,28,35]. LBD is a ''de facto standard'' technique until a complete intraoperative diagnostic system is established.…”

LINE-1 and Alu Methylation Patterns in Lymph Node Metastases of Head and Neck Cancers

“…These actions can cause inaccurate tumor staging and lead to improper therapeutic management (Gu, 2002;Coello, 2004;Imoto, 2006;Broglie, 2011;Rahbari, 2012). Many studies have developed methods for detecting occult tumor cells or DNA, including serial section staining, immunohistochemistry, PCR and RT-PCR-based methods (Tsavellas, 2001;Riethdorf, 2008;Wada, 2008;Shimizu, 2012); however, their conclusions have been less satisfactory. In this study, we aimed to determine whether LINE-1 and Alu methylation measurements are effective biomarkers for the detection of occult metastatic HNSCC and whether these measurements can provide a supportive method for diagnosing nodal metastases from HNSCC as an alternative to routine pathological examination.…”

Sentinel Node and Mechanism of Lymphatic Metastasis

“…11,12) Even when false-negative cases occur due to technical problems of SN identification and overlooked findings of intraoperative rapid cytodiagnosis, actual lymph node metastasis is present in the SN basin and can be frequently identified. 13) In patients with clear SN metastasis, since the possibility that metastasis is also present outside the SN basin cannot be excluded, standard resection and dissection are necessary at present.…”