, Volume 13, Issue 2, pp 167–172 | Cite as

Clinical significance of serum autoantibodies against Ras-like GTPases, RalA, in patients with esophageal squamous cell carcinoma

  • Tatsuki Nanami
  • Hideaki ShimadaEmail author
  • Satoshi Yajima
  • Yoko Oshima
  • Kazuyuki Matsushita
  • Fumio Nomura
  • Matsuo Nagata
  • Masatoshi Tagawa
  • Seiko Otsuka
  • Akiko Kuwajima
  • Hironori Kaneko
Original Article



The Ras-like GTPases, RalA and RalB are members of the Ras superfamily of small GTPases. Aberrant activation of Ral is a major cause of human tumorigenesis induced by oncogenic Ras. Serum anti-RalA antibodies are induced in esophageal carcinoma patients. However, detailed comparisons of their pathological characteristics are unavailable, and conventional serum markers have not been well evaluated.


Serum samples of 171 patients with esophageal squamous cell carcinoma and 73 healthy individuals were analyzed using specifically developed ELISA system for serum anti-RalA antibodies. A cut-off optical density value was fixed at 0.255 (the control mean + 2 SD). Clinicopathological characteristics and positive rates of conventional tumor markers were evaluated for seropositive patients.


Overall positive rate for serum anti-RalA antibodies was 18 %, which gradually increased with the tumor stages. Although the positive rate for serum anti-RalA antibodies was comparable with that of carcinoembryonic antigen (24 %) and CYFRA21-1 (21 %), it was lower than the rate for serum p53 antibodies (31 %) and squamous cell carcinoma antigen (37 %). Although serum anti-RalA antibodies were not associated with other serum markers, it was inversely associated with serum p53 antibodies. No clear association was observed between serum anti-RalA antibodies and RalA immunoreactivity.


Presence of serum anti-RalA antibodies is associated with tumor stages, but not with conventional tumor markers. Serum anti-RalA antibodies may be candidate serum markers in combination with other serum markers for esophageal squamous cell carcinoma.


RalA Serum autoantibody Tumor marker Esophageal cancer ELISA 



Carcinoembryonic antigen


Squamous cell carcinoma


Optical density


Serum RalA antibodies


Serum p53 antibodies



This research was partly supported by Grant-in-Aid for Scientific Research (Nos. 24591961 and 21591717) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Compliance with ethical standards

Ethical Statement

This study was approved by the institutional review boards of Chiba Cancer Center (#21-26) and Toho University School of Medicine (#22-112, #22-047). Additional informed consent was obtained from all patients for whom identifying information is included in this article.

Conflict of Interest

Hideaki Shimada received research grants and technical lecture fees form Medical & Biological Laboratories Co., Ltd., Nagoya, Japan. Akiko Kuwajima is an employee of Medical & Biological Laboratories Co., Ltd., Nagoya, Japan. The other authors have no conflict of interest.


  1. 1.
    Shirakawa R, Horiuchi H. Ral GTPases: crucial mediators of exocytosis and tumorigenesis. J Biochem. 2015;. doi: 10.1093/jb/mvv029.Google Scholar
  2. 2.
    Schubbert S, Shannon K, Bollag G. Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer. 2007;7:295–308.CrossRefPubMedGoogle Scholar
  3. 3.
    Bodemann BO, White MA. Ral GTPases and cancer: linchpin support of the tumorigenic platform. Nat Rev Cancer. 2008;8:133–40.CrossRefPubMedGoogle Scholar
  4. 4.
    Hamad NM, Elconin JH, Karnoub AE, et al. Distinct requirements for Ras oncogenesis in human versus mouse cells. Genes Dev. 2002;16:2045–57.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Oxford G, Owens CR, Titus BJ, et al. RalA and RalB: antagonistic relatives in cancer cell migration. Cancer Res. 2005;65:7111–20.CrossRefPubMedGoogle Scholar
  6. 6.
    Vigil D, Martin TD, Williams F, et al. Aberrant overexpression of the Rgl2 Ral small GTPase-specific guanine nucleotide exchange factor promotes pancreatic cancer growth through Ral-dependent and Ral-independent mechanisms. J Biol Chem. 2010;285:34729–40.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Martin TD, Samuel JC, Routh ED, et al. Activation and involvement of Ral GTPases in colorectal cancer. Cancer Res. 2011;71:206–15.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Male H, Patel V, Jacob MA, et al. Inhibition of RalA signaling pathway in treatment of non-small cell lung cancer. Lung Cancer. 2012;77:252–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Guin S, Ru Y, Wynes MW, et al. Contributions of KRAS and RAL in non-small-cell lung cancer growth and progression. J Thorac Oncol. 2013;8:1492–501.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Smith SC, Baras AS, Owens CR, et al. Transcriptional signatures of Ral GTPase are associated with aggressive clinicopathologic characteristics in human cancer. Cancer Res. 2012;72:3480–91.CrossRefPubMedGoogle Scholar
  11. 11.
    Sowalsky AG, Alt-Holland A, Shamis Y, et al. RalA function in dermal fibroblasts is required for the progression of squamous cell carcinoma of the skin. Cancer Res. 2011;71:758–67.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Xu YW, Peng YH, Chen B, et al. Autoantibodies as potential biomarkers for the early detection of esophageal squamous cell carcinoma. Am J Gastroenterol. 2014;109:36–45.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Shimada H, Ochiai T, Nomura F. Japan p53 Antibody Research Group. Titration of serum p53 antibodies in 1,085 patients with various types of malignant tumors: a multiinstitutional analysis by the Japan p53 Antibody Research Group. Cancer. 2003;97:682–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Shimada H, Yajima S, Oshima Y, et al. Impact of serum biomarkers on esophageal squamous cell carcinoma. Esophagus. 2012;9:131–40.CrossRefGoogle Scholar
  15. 15.
    Oshima Y, Shimada H, Yajima S, et al. NY-ESO-1 autoantibody as a tumor-specific biomarker for esophageal cancer: screening in 1969 patients with various cancers. J Gastroenterol. 2015. doi: 10.1007/s00535-015-1078-8.
  16. 16.
    Wang K, Chen Y, Liu S, et al. Immunogenicity of Ra1A and its tissue-specific expression in hepatocellular carcinoma. Int J Immunopathol Pharmacol. 2009;22:735–43.PubMedPubMedCentralGoogle Scholar
  17. 17.
    Qin JJ, Wang XR, Wang P, et al. Mini-array of multiple tumor-associated antigens (TAAs) in the immunodiagnosis of esophageal cancer. Asian Pac J Cancer Prev. 2014;15:2635–40.CrossRefPubMedGoogle Scholar
  18. 18.
    Shimada H, Nabeya Y, Okazumi S, et al. Prediction of survival with squamous cell carcinoma antigen in patients with resectable esophageal squamous cell carcinoma. Surgery. 2003;133:486–94.CrossRefPubMedGoogle Scholar
  19. 19.
    Shimada H, Nabeya Y, Okazumi S, et al. Prognostic significance of CYFRA 21-1 in patients with esophageal squamous cell carcinoma. J Am Coll Surg. 2003;196:573–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Tecleab A, Zhang X, Sebti SM. Ral GTPase down-regulation stabilizes and reactivates p53 to inhibit malignant transformation. J Biol Chem. 2014;289:31296–309.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Yan C, Liu D, Li L, et al. Discovery and characterization of small molecules that target the GTPase Ral. Nature. 2014;515:443–7.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Kidd AR 3rd, Snider JL, Martin TD, et al. Ras-related small GTPases RalA and RalB regulate cellular survival after ionizing radiation. Int J Radiat Oncol Biol Phys. 2010;78:205–12.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Japan Esophageal Society and Springer Japan 2015

Authors and Affiliations

  • Tatsuki Nanami
    • 1
  • Hideaki Shimada
    • 1
    Email author
  • Satoshi Yajima
    • 1
  • Yoko Oshima
    • 1
  • Kazuyuki Matsushita
    • 2
  • Fumio Nomura
    • 2
  • Matsuo Nagata
    • 3
  • Masatoshi Tagawa
    • 4
  • Seiko Otsuka
    • 1
  • Akiko Kuwajima
    • 5
  • Hironori Kaneko
    • 1
  1. 1.Department of Surgery, School of MedicineToho UniversityTokyoJapan
  2. 2.Department of Molecular DiagnosisChiba University, Graduate School of MedicineChibaJapan
  3. 3.Division of Gastroenterological SurgeryChiba Cancer CenterChibaJapan
  4. 4.Division of Pathology and Cell TherapyChiba Cancer CenterChibaJapan
  5. 5.Medical & Biological Laboratories Co., Ltd.NagoyaJapan

Personalised recommendations