Evaluation of <em>RASSF1A</em> methylation in the lysate of sentinel lymph nodes for detecting breast cancer metastasis: A diagnostic accuracy study

Park,Sung Ae; Masunaga,Nanae; Kagara,Naofumi; Ohi,Yasuyo; Gondo,Naomi; Abe,Kaori; Yoshinami,Tetsuhiro; Sota,Yoshiaki; Miyake,Tomohiro; Tanei,Tomonori; Shimoda,Masafumi; Sagara,Yasuaki; Shimazu,Kenzo

doi:10.3892/ol.2023.14063

Evaluation of RASSF1A methylation in the lysate of sentinel lymph nodes for detecting breast cancer metastasis: A diagnostic accuracy study

Authors:
- Sung Ae Park
- Nanae Masunaga
- Naofumi Kagara
- Yasuyo Ohi
- Naomi Gondo
- Kaori Abe
- Tetsuhiro Yoshinami
- Yoshiaki Sota
- Tomohiro Miyake
- Tomonori Tanei
- Masafumi Shimoda
- Yasuaki Sagara
- Kenzo Shimazu
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Affiliations: Department of Breast and Endocrine Surgery, Osaka University Graduate School of Medicine, Suita, Osaka 565‑0871, Japan, Department of Breast Surgery, Osaka General Medical Center, Osaka 558‑8558, Japan, Department of Breast Surgery, Hakuaikai Sagara Hospital, Kagoshima 892‑0833, Japan
Published online on: September 21, 2023 https://doi.org/10.3892/ol.2023.14063
Article Number: 475
Copyright: © Park et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The restriction enzyme‑based digital methylation‑specific polymerase chain reaction (RE‑dMSP) assay is useful for diagnosing sentinel lymph node (SN) metastasis in patients with breast cancer, by detecting tumor‑derived methylated Ras association domain‑containing protein 1 (RASSF1A). In addition, this assay has high concordance (95.0%) with one‑step nucleic acid amplification (OSNA). The present study aimed to perform RE‑dMSP using OSNA lysate from more patients and to re‑evaluate its clinical usage. Overall, 418 SNs from 347 patients were evaluated using both OSNA and RE‑dMSP. The concordance rate was 83.3% (348/418). RASSF1A methylation of the primary tumors was negative in 36 patients. When these patients were excluded, the concordance rate improved to 88.2% (330/374). Of the 79 OSNA‑negative cases, 19 were RE‑dMSP‑positive, although all were positive for cytokeratin 19 expression in the primary tumor, suggesting that RE‑dMSP can detect tumor‑derived DNA with a higher sensitivity. The percent of methylated reference of the breast tumors showed a wide variety in the 16 OSNA‑positive/RE‑dMSP‑negative cases, and such variability of methylation could have affected the results in these patients. In conclusion, although RE‑dMSP can diagnose SN metastasis with high sensitivity and accuracy, and can be a supplementary tool to OSNA in breast cancer, RE‑dMSP showed certain discordance with OSNA and critically depended on the absence or heterogeneity of DNA methylation in breast tumors. Further research is expected to develop an assay targeting other DNA alterations, such as mutations.

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