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Identification and Validation of DNA Methylation Markers to Predict Lymph Node Metastasis of Esophageal Squamous Cell Carcinomas

  • Translational Research and Biomarkers
  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

Abstract

Background

The presence of lymph node metastasis in esophageal squamous cell carcinoma (ESCC) patients is a critical factor for decision of treatment strategy. However, there have been no molecular markers to assess lymph node metastasis. In this study, we aimed to identify CpG islands (CGIs) whose DNA methylation statuses are associated with the presence of lymph node metastasis.

Materials and Methods

A total of 96 ESCCs were divided into a screening set (n = 48) and a validation set (n = 48). Genome-wide methylation analysis was performed by methylated DNA immunoprecipitation-CGI microarray analysis. Methylation levels were analyzed by quantitative methylation-specific PCR (qMSP).

Results

Genome-wide methylation analysis identified 25 CGIs differentially methylated between 8 ESCCs with lymph node metastasis and 4 without. In the screening set, 7 CGIs had significantly different methylation levels (P < 0.05) between the ESCCs with and without lymph node metastasis, and cut-off methylation levels for these CGIs were determined. The validation set was analyzed with the prefixed cut-offs, and methylation statuses of 2 CGIs in the vicinities of PAX6 and ENST00000363328 were validated to be associated with the presence of lymph node metastasis. Using these 2 markers, the presence was predicted with a sensitivity of 93% and specificity of 57%. In addition, the methylation statuses of the 2 CGIs were significantly associated with disease-free survival (P = 0.006).

Conclusions

Methylation statuses of these 2 CGIs were significantly associated with the presence of lymph node metastasis of ESCCs. These CGIs are promising markers to predict the presence of lymph node metastases.

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Acknowledgment

The authors are grateful to Dr. Taichi Shimazu for his advice in statistical analysis. This study was supported by Grants-in-Aid for Cancer Research and for the Third-Term Comprehensive Cancer Control Strategy from the Ministry of Health, Labour and Welfare, Japan. K.G. and Y.M. are recipients of Research Resident Fellowships from the Foundation for Promotion of Cancer Research.

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Authors and Affiliations

  1. Carcinogenesis Division, National Cancer Center Research Institute, Tokyo, Japan

    Ken Gyobu MD, Satoshi Yamashita PhD, Yasunori Matsuda MD, Tohru Niwa PhD, Daiji Oka MD & Toshikazu Ushijima MD, PhD

  2. Department of Gastrointestinal Surgery, Graduate School of Medicine, Osaka City University, Osaka, Japan

    Ken Gyobu MD, Yasunori Matsuda MD, Harushi Osugi MD, PhD & Shigeru Lee MD

  3. Esophageal Surgery Division, National Cancer Center Hospital, Tokyo, Japan

    Hiroyasu Igaki MD, PhD

  4. Pathology of Clinical Laboratory Division, National Cancer Center Hospital, Tokyo, Japan

    Ryoji Kushima MD, PhD

  5. Department of Cardiovascular Surgery, Graduate School of Medicine, Osaka City University, Osaka, Japan

    Shigefumi Suehiro MD, PhD

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  1. Ken Gyobu MD
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  2. Satoshi Yamashita PhD
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  3. Yasunori Matsuda MD
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  4. Hiroyasu Igaki MD, PhD
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  11. Toshikazu Ushijima MD, PhD
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Correspondence to Toshikazu Ushijima MD, PhD.

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Gyobu, K., Yamashita, S., Matsuda, Y. et al. Identification and Validation of DNA Methylation Markers to Predict Lymph Node Metastasis of Esophageal Squamous Cell Carcinomas. Ann Surg Oncol 18, 1185–1194 (2011). https://doi.org/10.1245/s10434-010-1393-5

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  • DOI: https://doi.org/10.1245/s10434-010-1393-5

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