Annals of Surgical Oncology

, Volume 22, Issue 3, pp 980–986 | Cite as

A New Diagnostic Method for Rapid Detection of Lymph Node Metastases Using a One-Step Nucleic Acid Amplification (OSNA) Assay in Endometrial Cancer

  • Tomoyuki Nagai
  • Hitoshi Niikura
  • Satoshi Okamoto
  • Kadzuki Nakabayashi
  • Maki Matoda
  • Hiroki Utsunomiya
  • Satoru Nagase
  • Mika Watanabe
  • Nobuhiro Takeshima
  • Nobuo Yaegashi
Gynecologic Oncology

Abstract

Background

To improve lymph node (LN) metastasis identification for patients with endometrial cancer (EC), this study assessed the usefulness of molecular biologic techniques using a one-step nucleic acid amplification (OSNA) assay.

Methods

Using quantitative reverse transcription polymerase chain reaction (qRT-PCR), an optimal mRNA marker was selected, and its expression was compared between histopathologically positive and negative LNs using an OSNA assay. The authors determined copy number cutoff values and evaluated the diagnostic performance of this OSNA assay using sentinel lymph nodes (SLNs). They also investigated whether an OSNA assay could detect LN metastases with sensitivity and specificity equivalent to the 2-mm-interval histopathology method.

Results

For analysis of EC samples, cytokeratin 19 (CK19) was selected as a useful mRNA marker for the OSNA assay. When the cutoff value was set at 250 copies (using 215 LNs from 70 patients), an OSNA assay using CK19 mRNA had a sensitivity of 93.3 %, a specificity of 99.5 %, and a concordance rate of 99.1 %. For performance evaluations using SLNs (120 histopathologically negative LNs and 17 histopathologically positive LNs from 35 patients), a OSNA assay using CK19 mRNA had a sensitivity of 82.4 %, a specificity of 99.2 %, a positive predictive value of 93.3 %, and a concordance rate of 97.1 %. Thus, an OSNA assay using CK19 mRNA provided results equivalent to those with the 2-mm-interval histopathology method.

Conclusions

The study data demonstrated that an OSNA assay using CK19 mRNA was applicable for detecting LN metastases in EC. Combined analysis using an OSNA assay and SLNs may improve individualized treatments according to LN metastatic status.

Keywords

Sentinel Lymph Node Endometrial Cancer National Comprehensive Cancer Network National Comprehensive Cancer Network Sentinel Lymph Node Metastasis 

Notes

Acknowledgment

The authors thank Ms. Emi Endo, Ms. Aya Miyabe, Mr. Hiroyuki Shimizu, and Mr. Kengo Goto for their substantial support. This study was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science and Culture, Japan; a Grant-in-Aid for Scientific Research (B) and (C); a Grant-in-Aid for Young Scientists (B); a Grant-in-Aid for Exploratory Research, from the Ministry of Education, Science, Sports and Culture, Japan; a Grant-in-Aid from the Ministry of Health, Labor and Welfare, Japan; the Global COE Program Special Research Grant (Tohoku University) from the Ministry of Education Science, Sports and Culture, Japan; Kurokawa Cancer Research Foundation, and the Uehara Memorial Foundation.

Conflict of interest

There are no conflicts of interest.

References

  1. 1.
    ASTEC study group; Kitchener H, et al. Efficacy of systematic pelvic lymphadenectomy in endometrial cancer (MRC ASTEC trial): a randomised study. Lancet. 2009:373:125–36.PubMedCrossRefGoogle Scholar
  2. 2.
    Benedetti-Panici P, Basile S, Maneschi F, Alberto-Lissoni A, Signorelli M, Scambia G, et al. Systematic pelvic lymphadenectomy vs. no lymphadenectomy in early-stage endometrial carcinoma: randomized clinical trial. J Natal Cancer Inst. 2008;100:1707–16.CrossRefGoogle Scholar
  3. 3.
    To-do Y, Kato H, Kaneohe M, Watery H, Tread M, Samurai N Survival effect of para-aortic lymphadenectomy in endometrial cancer (SEPAL study): a retrospective cohort analysis. Lancet. 2010;375:1165–72.CrossRefGoogle Scholar
  4. 4.
    NCCN Clinical Practice Guidelines in Oncology Uterine neoplasms, version 1, 2015.Google Scholar
  5. 5.
    Altgassen C, Muller N, Hornemann A, Kavallaris A, Hornung D, Diedrich K, et al. Immunohistochemical workup of sentinel nodes in endometrial cancer improves diagnostic accuracy. Gynecol Oncol. 114:284–7.Google Scholar
  6. 6.
    Amezuca CA, Macdonald HR, Lum CA, Yi W, Muderspach LI, Roman LD, et al. Endometrial cancer patients have a significant risk of harboring isolated tumor cells in histologically negative lymph nodes. Int J Gynecol Cancer. 2006;16:1336–41.CrossRefGoogle Scholar
  7. 7.
    Niikura H, Okamoto S, Yoshinaga K, Nagase S, Takano T, Ito K, et al. Detection of micrometastases in the sentinel lymph nodes patients with endometrial cancer. Gynecol Oncol. 2007;105:683–6.PubMedCrossRefGoogle Scholar
  8. 8.
    Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000;28:E63.PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Tsujimoto M, Nakabayashi K, Yoshidome K, Kaneko T, Iwase T, Akiyama F, et al. One-step nucleic acid amplification for intraoperative detection of lymph node metastasis in breast cancer patients. Clin Cancer Res. 2007;13:4807–16.PubMedCrossRefGoogle Scholar
  10. 10.
    Tamaki Y, Sato N, Homma K, Takabatake D, Nishimura R, Tsujimoto M, et al. Routine clinical use of the one-step nucleic acid amplification assay for detection of sentinel lymph node metastases in breast cancer patients. Cancer. 2012;15:3477–83.CrossRefGoogle Scholar
  11. 11.
    Osako T, Iwase T, Kimura K, Yamashita K, Hori R, Yanagisawa A, et al. Intraoperative molecular assay for sentinel lymph node metastases in early stage breast cancer. Cancer. 2011;19:4365–74.CrossRefGoogle Scholar
  12. 12.
    Croner RS, Schellerer V, Demund H, Shildberg C, Papadopulos T, Nachberger E, et al. One-step nucleic acid amplification (OSNA): a new method for lymph node staging in colorectal carcinomas. J Transl Med. 2010;8:83–8.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Yaguchi Y, Sugasawa H, Tsujimoto H, Takata H, Nakabayashi K, Ichikura T, et al. One-step nucleic acid amplification (OSNA) for the application of sentinel node concept in gastric cancer. Ann Surg Oncol. 2011;18:2289–96.PubMedCrossRefGoogle Scholar
  14. 14.
    Yamamoto N, Daito M, Hiyama K, Ding J, Nakabayashi K, Otomo Y, et al. An optimal mRNA marker for OSNA based lymph node metastasis detection in colorectal cancer patients. Jpn J Clin Oncol. 2013;43:264–70.PubMedCrossRefGoogle Scholar
  15. 15.
    Okamoto S, Niikura H, Nakabayashi K, Hiyama K, Matoda M, Takeshima N, et al. Detection of sentinel lymph node metastases in cervical cancer: assessment of KRT19 mRNA in the one-step nucleic acid amplification (OSNA) method. Gynecol Oncol. 2013;130:530–6.PubMedCrossRefGoogle Scholar
  16. 16.
    Fishman A, Klein A, Zemer R, Sc M, Zimlichman S, Bernheim J, et al. Detection of micrometastasis by cytokeratin-20 (reverse transcription polymerase chain reaction) in lymph nodes of patients with endometrial cancer. Gynecol Oncol. 2000;77:399–404.PubMedCrossRefGoogle Scholar
  17. 17.
    Xin-Qiang Ji, Sato H, Tanaka H, Konishi Y, Fujimoto T, Takahashi O, et al. Real-time quantitative RT-PCR detection of disseminated endometrial tumor cells in peripheral blood and lymph nodes using the LightCycler system. Gynecol Oncol. 2006;100:355–60.CrossRefGoogle Scholar
  18. 18.
    Niikura H, Kaiho-Sakuma M, Tokunaga H, Toyoshima M, Utsunomiya H, Nagase S, et al. Tracer injection sites and combinations for sentinel lymph node detection in patients with endometrial cancer. Gynecol Oncol. 2013:131:299–303.PubMedCrossRefGoogle Scholar
  19. 19.
    Stewart C, Crook M.L, Lacey J, Louwen K, et al. Cytokeratin 19 expression in normal endometrium and in low-grade endometrioid adenocarcinoma of the endometrium. Int J Gynecol Pathol. 2011;30:484–91.PubMedCrossRefGoogle Scholar
  20. 20.
    Tamaki Y, Akiyama F, Iwase T, Kaneko T, Tsuda H, Sato K, et al. Molecular detection of lymph node metastases in breast cancer patients: results of a multicenter trial using the one-step nucleic acid amplification assay. Clin Cancer Res. 2009;15:2879–84.PubMedCrossRefGoogle Scholar
  21. 21.
    Yamamoto H, Sekimoto M, Oya M, Yamamoto N, Konishi F, Sasaki J, et al. OSNA-based novel molecular testing for lymph node metastases in colorectal cancer patients: results from a multicenter clinical performance study in Japan. Ann Surg Oncol. 2011;18:1891–8.PubMedCrossRefGoogle Scholar
  22. 22.
    Kumagai K, Yamamoto N, Miyashiro I, Tomita Y, Katai H, Kushima R, et al. Multicenter study evaluating the clinical performance of the OSNA assay for the molecular detection of lymph node metastases in gastric cancer patients. Gastric Cancer. 2014;17:273–80.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Niikura H, Okamura C, Utsunomiya H, Yoshinaga K, Akahira J, Ito K, et al. Sentinel lymph node detection in patients with endometrial cancer. Gynecol Oncol. 2004;92:669–74.PubMedCrossRefGoogle Scholar
  24. 24.
    Ballester M, Dubernard G, Lecuru F, Heitz D, Mathevet P, Marret H, et al. Detection rate and diagnostic accuracy of sentinel-node biopsy in early-stage endometrial cancer: a prospective multicentre study (SENTI-ENDO). Lancet Oncol. 2011;12:469–76.PubMedCrossRefGoogle Scholar
  25. 25.
    Abu-Rustum NR, Gomez JD, Alektiar KM, Soslow RA, Hensley ML, Leitao Jr MM, et al. The incidence of isolated paraaortic nodal metastasis in surgically staged endometrial cancer patients with negative pelvic lymph nodes. Gynecol Oncol. 2009;115:236–8.PubMedCrossRefGoogle Scholar

Copyright information

© Society of Surgical Oncology 2014

Authors and Affiliations

  • Tomoyuki Nagai
    • 1
  • Hitoshi Niikura
    • 1
  • Satoshi Okamoto
    • 1
  • Kadzuki Nakabayashi
    • 2
  • Maki Matoda
    • 3
  • Hiroki Utsunomiya
    • 1
  • Satoru Nagase
    • 1
  • Mika Watanabe
    • 4
  • Nobuhiro Takeshima
    • 3
  • Nobuo Yaegashi
    • 1
  1. 1.Department of Obstetrics and GynecologyTohoku University School of MedicineSendaiJapan
  2. 2.Department of Product Development of LS Business UnitSysmex CorporationKobeJapan
  3. 3.Department of GynecologyCancer Institute HospitalTokyoJapan
  4. 4.Department of PathologyTohoku University HospitalSendaiJapan

Personalised recommendations