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Flow Cytometric Quantification of Intraperitoneal Free Tumor Cells is a Useful Biomarker in Gastric Cancer Patients with Peritoneal Metastasis

  • Gastrointestinal Oncology
  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

Abstract

Background

The frequency of intraperitoneal free tumor cells (IPTC) is considered to reflect the severity of peritoneal metastasis (PM). We quantified the relative number of IPTC against leukocytes in peritoneal fluid and evaluated its clinical relevance in gastric cancer (GC) patients, particularly those with PM.

Methods

Cells recovered from ascites or peritoneal lavage fluid were immunostained with monoclonal antibodies (mAb) to CD45 and CD326 (EpCAM). Using flow cytometry (FACS), CD326(+) and CD45(+) cells were classified as either tumor cells (T) or leukocytes (L) and the T/L ratio (TLR) was calculated in a total of 506 samples obtained from 300 patients with GC and 33 patients with liver cirrhosis (LC).

Results

Median (M) of the TLR of the initial samples obtained from 199 patients with PM(+) GC was 1.32 % (0–1,868.44 %), which was significantly higher than that in patients with PM(−) GC (M = 0 %, 0–0.35 %; n = 101) or LC (M = 0 %, 0–0.031 %; n = 33). In 104 PM(+) patients who received combination chemotherapy including intraperitoneal paclitaxel, the TLR was repeatedly measured in peritoneal fluid obtained from the port. In these patients, the TLR showed a strong correlation with clinical features as well as cytological findings and carcinoembryonic antigen messenger RNA status. Finally, the median survival time of the 11 patients with initial TLR > 10 % was significantly shorter than that of the 52 patients with TLR < 10 % (271 vs. 627 days; p = 0.0002).

Conclusion

The TLR excellently reflected tumor burden in the peritoneal cavity, and could be a reliable biomarker to determine the outcome, as well as the effectiveness, of chemotherapy in patients with PM(+) GC.

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References

  1. Nakamura K, Ueyama T, Yao T, et al. Pathology and prognosis of gastric carcinoma: findings in 10,000 patients who underwent primary gastrectomy. Cancer. 1992;70:1030–1037.

    Article  CAS  PubMed  Google Scholar 

  2. Isobe Y, Nashimoto A, Akazawa K, et al. Gastric cancer treatment in Japan: 2008 annual report of the JGCA nationwide registry. Gastric Cancer. 2011;14:301–316.

    Article  PubMed Central  PubMed  Google Scholar 

  3. Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.

    Article  PubMed  Google Scholar 

  4. Koga S, Kaibara N, Iitsuka Y, et al. Prognostic significance of intraperitoneal free cancer cells in gastric cancer patients. J Cancer Res Clin Oncol. 1984;108:236–238.

    Article  CAS  PubMed  Google Scholar 

  5. Sodek KL, Murphy KJ, Brown TJ, Ringuette MJ. Cell-cell and cell-matrix dynamics in intraperitoneal cancer metastasis. Cancer Metastasis Rev. 2012;31:397–414.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Warshaw AL. Implications of peritoneal cytology for staging of early pancreatic cancer. Am J Surg. 1991;161:26–29; discussion 29–30.

    Article  CAS  PubMed  Google Scholar 

  7. Ziselman EM, Harkavy SE, Hogan M, et al. Peritoneal washing cytology. Uses and diagnostic criteria in gynecologic neoplasms. Acta Cytol. 1984;28:105–110.

    CAS  PubMed  Google Scholar 

  8. Hayes N, Wayman J, Wadehra V, et al. Peritoneal cytology in the surgical evaluation of gastric carcinoma. Br J Cancer. 1999;79:520–524.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Vogel P, Ruschoff J, Kummel S, et al. Prognostic value of microscopic peritoneal dissemination: comparison between colon and gastric cancer. Dis Colon Rectum. 2000;43:92–100.

    Article  CAS  PubMed  Google Scholar 

  10. Horattas MC, Evasovich MR, Topham N. Colorectal carcinoma and the relationship of peritoneal cytology. Am J Surg. 1997;174:334–337; discussion 337–338.

    Article  CAS  PubMed  Google Scholar 

  11. Bonenkamp JJ, Songun I, Hermans J, van de Velde CJ. Prognostic value of positive cytology findings from abdominal washings in patients with gastric cancer. Br J Surg. 1996;83:672–674.

    Article  CAS  PubMed  Google Scholar 

  12. Bando E, Yonemura Y, Takeshita Y, et al. Intraoperative lavage for cytological examination in 1,297 patients with gastric carcinoma. Am J Surg. 1999;178:256–262.

    Article  CAS  PubMed  Google Scholar 

  13. Leake PA, Cardoso R, Seevaratnam R, et al. A systematic review of the accuracy and utility of peritoneal cytology in patients with gastric cancer. Gastric Cancer. 2012;15 Suppl 1:S27–S37.

    Article  PubMed  Google Scholar 

  14. Konishi M, Kinoshita T, Nakagohri T, et al. Prognostic value of cytologic examination of peritoneal washings in pancreatic cancer. Arch Surg. 2002;137:475–480.

    Article  PubMed  Google Scholar 

  15. Lorenzen S, Panzram B, Rosenberg R, et al. Prognostic significance of free peritoneal tumor cells in the peritoneal cavity before and after neoadjuvant chemotherapy in patients with gastric carcinoma undergoing potentially curative resection. Ann Surg Oncol. 2010;17:2733–2739.

    Article  PubMed  Google Scholar 

  16. Katsuragi K, Yashiro M, Sawada T, et al. Prognostic impact of PCR-based identification of isolated tumour cells in the peritoneal lavage fluid of gastric cancer patients who underwent a curative R0 resection. Br J Cancer. 2007;97:550–556.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Kodera Y, Nakanishi H, Ito S, et al. Quantitative detection of disseminated free cancer cells in peritoneal washes with real-time reverse transcriptase-polymerase chain reaction: a sensitive predictor of outcome for patients with gastric carcinoma. Ann Surg. 2002;235:499–506.

    Article  PubMed Central  PubMed  Google Scholar 

  18. Rossi Del Monte S, Ranieri D, Mazzetta F, et al. Free peritoneal tumor cells detection in gastric and colorectal cancer patients. J Surg Oncol. 2012;106:17–23.

    Article  CAS  PubMed  Google Scholar 

  19. Dalal KM, Woo Y, Kelly K, et al. Detection of micrometastases in peritoneal washings of gastric cancer patients by the reverse transcriptase polymerase chain reaction. Gastric Cancer. 2008;11:206–213.

    Article  CAS  PubMed  Google Scholar 

  20. Kowalewska M, Chechlinska M, Nowak R. Carcinoembryonic antigen and cytokeratin 20 in peritoneal cells of cancer patients: are we aware of what we are detecting by mRNA examination? Br J Cancer. 2008;98:512–513; author reply 514.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Kitayama J, Emoto S, Yamaguchi H, et al. Flow cytometric quantification of intraperitoneal free tumor cells in patients with peritoneal metastasis. Cytometry B Clin Cytom. 2014;86:56–62.

    Article  PubMed  Google Scholar 

  22. Ishigami H, Kitayama J, Kaisaki S, et al. Phase II study of weekly intravenous and intraperitoneal paclitaxel combined with S-1 for advanced gastric cancer with peritoneal metastasis. Ann Oncol. 2010;21:67–70.

    Article  CAS  PubMed  Google Scholar 

  23. Yamaguchi H, Kitayama J, Ishigami H, et al. A phase 2 trial of intravenous and intraperitoneal paclitaxel combined with S-1 for treatment of gastric cancer with macroscopic peritoneal metastasis. Cancer. 2013;119:3354–3358.

    Article  CAS  PubMed  Google Scholar 

  24. Benevolo M, Mottolese M, Cosimelli M, et al. Diagnostic and prognostic value of peritoneal immunocytology in gastric cancer. J Clin Oncol. 1998;16:3406–3411.

    CAS  PubMed  Google Scholar 

  25. Vogel P, Ruschoff J, Kummel S, et al. Immunocytology improves prognostic impact of peritoneal tumour cell detection compared to conventional cytology in gastric cancer. Eur J Surg Oncol. 1999;25:515–519.

    Article  CAS  PubMed  Google Scholar 

  26. Went PT, Lugli A, Meier S, et al. Frequent EpCam protein expression in human carcinomas. Hum Pathol. 2004;35:122–128.

    Article  CAS  PubMed  Google Scholar 

  27. Baeuerle PA, Gires O. EpCAM (CD326) finding its role in cancer. Br J Cancer. 2007;96:417–423.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Patriarca C, Macchi RM, Marschner AK, Mellstedt H. Epithelial cell adhesion molecule expression (CD326) in cancer: a short review. Cancer Treat Rev. 2012;38:68–75.

    Article  CAS  PubMed  Google Scholar 

  29. Burges A, Wimberger P, Kumper C, et al. Effective relief of malignant ascites in patients with advanced ovarian cancer by a trifunctional anti-EpCAM x anti-CD3 antibody: a phase I/II study. Clin Cancer Res. 2007;13:3899–3905.

    Article  CAS  PubMed  Google Scholar 

  30. Heiss MM, Murawa P, Koralewski P, et al. The trifunctional antibody catumaxomab for the treatment of malignant ascites due to epithelial cancer: results of a prospective randomized phase II/III trial. Int J Cancer. 2010;127:2209–2221.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  31. Miyashiro I, Takachi K, Doki Y, et al. When is curative gastrectomy justified for gastric cancer with positive peritoneal lavage cytology but negative macroscopic peritoneal implant? World J Surg. 2005;29:1131–1134.

    Article  PubMed  Google Scholar 

  32. Iitsuka Y, Shiota S, Matsui T, et al. Relationship between the cytologic characteristics of intraperitoneal free cancer cells and the prognosis in patients with gastric cancer. Acta Cytol. 1990;34:437–442.

    CAS  PubMed  Google Scholar 

  33. Kitayama J, Ishigami H, Yamaguchi H, et al. Salvage gastrectomy after intravenous and intraperitoneal paclitaxel (PTX) administration with oral S-1 for peritoneal dissemination of advanced gastric cancer with malignant ascites. Ann Surg Oncol. 2014;21(2):539–546.

    Article  PubMed  Google Scholar 

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Acknowledgment

We thank Ms. Chieko Uchikawa, Ikuko Nieda, and Yu Saito for their excellent technical and clerical support. This study was financially supported by the Ministry of Health, Labor and Welfare of Japan. This work was funded by the Ministry of Education, Culture, Sports, Science and Technology of Japan, and the Ministry of Health, Labor and Welfare of Japan, and by a Grant-in-aid of the Public Trust Surgery Research Fund, Tokyo, Japan.

Disclosure

Joji Kitayama, Shigenobu Emoto, Hironori Yamaguchi, Hironori Ishigami, Haruna Onoyama, Hiroharu Yamashita, Yasuyuki Seto, Keisuke Matsuzaki, and Toshiaki Watanabe have no financial disclosures.

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Authors and Affiliations

  1. Department of Surgical Oncology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Joji Kitayama MD, Shigenobu Emoto MD, Hironori Yamaguchi MD, Hironori Ishigami MD & Toshiaki Watanabe MD

  2. Department of Gastrointestinal Surgery, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Haruna Onoyama MD, Hiroharu Yamashita MD & Yasuyuki Seto MD

  3. Kanamecho Hospital, Tokyo, Japan

    Keisuke Matsuzaki MD

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  1. Joji Kitayama MD
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Correspondence to Joji Kitayama MD.

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Kitayama, J., Emoto, S., Yamaguchi, H. et al. Flow Cytometric Quantification of Intraperitoneal Free Tumor Cells is a Useful Biomarker in Gastric Cancer Patients with Peritoneal Metastasis . Ann Surg Oncol 22, 2336–2342 (2015). https://doi.org/10.1245/s10434-014-4238-9

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  • DOI: https://doi.org/10.1245/s10434-014-4238-9

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