Journal of Gastroenterology

, Volume 45, Issue 12, pp 1201–1211

Dermokine as a novel biomarker for early-stage colorectal cancer

  • Tomoyuki Tagi
  • Takeshi Matsui
  • Shojiro Kikuchi
  • Sachi Hoshi
  • Toshiya Ochiai
  • Yukihito Kokuba
  • Yoko Kinoshita-Ida
  • Fumie Kisumi-Hayashi
  • Koji Morimoto
  • Toshio Imai
  • Issei Imoto
  • Johji Inazawa
  • Eigo Otsuji
Original Article—Alimentary Tract

DOI: 10.1007/s00535-010-0279-4

Cite this article as:
Tagi, T., Matsui, T., Kikuchi, S. et al. J Gastroenterol (2010) 45: 1201. doi:10.1007/s00535-010-0279-4

Abstract

Background

Colorectal cancer is a common disease that is usually detected at an advanced stage, because early-stage cancer is mostly asymptomatic and appropriate serologic biomarkers have not been established. We have previously identified dermokine (DK) as a peptide secreted by keratinocytes and we found that DK-β/γ was expressed in colorectal tumors. Therefore, we focused on DK-β/γ as a new candidate diagnostic serum marker for early colorectal cancer.

Methods

DK-β/γ expression in human colorectal cancer cell lines and tissues was assessed by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. We established an experimental enzyme-linked immunosorbent assay (ELISA) to detect DK-β/γ in the serum of colorectal cancer patients, and we compared the sensitivities of common diagnostic markers, carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, and serum p53 antibody (S-p53).

Results

Immunohistochemical staining of colon tumor tissue with anti-DK monoclonal antibody (mAb) revealed that DK-β/γ was more commonly expressed in the early stages of colorectal cancer (Tis–T1; i.e., cancer in situ, intraepithelial or invasion of lamina propria [Tis]; tumor invades the submucosa [T1]) than in late-stage tumors (T2–T4; i.e., tumor invades the muscularis propria [T2]; tumor invades through the muscularis propria into the subserosa, or into the nonperitonealized pericolic or perirectal tissues [T3]; tumor directly invades other organs or structures and/or perforates visceral peritoneum [T4]). Serum DK-β/γ levels were determined in 130 patients with colorectal cancer and 25 healthy volunteers. Serum DK-β/γ was detected in 33.3% of patients with early colorectal cancer (Tis–T1), which was higher than the rates for S-p53 (24.2%), CEA (9.1%), and CA19-9 (0%). The serum DK-β/γ test was complementary to the other marker tests. Therefore, when the combined four-marker test (DK/CEA/CA19-9/S-p53) was carried out, the diagnostic sensitivity for Tis and T1 tumors reached 60.6%.

Conclusions

Serum DK-β/γ is the most promising of the existing tumor biomarkers for the diagnosis of early-stage colorectal cancer.

Keywords

Serum markerDermokineColorectal cancerEarly stage

Abbreviations

DK

Dermokine

CEA

Carcinoembryonic antigen

CA19-9

Carbohydrate antigen 19-9

S-p53

Serum p53 antibody

ELISA

Enzyme-linked immunosorbent assay

SSE

Stratified squamous epithelium

Tis

Cancer in situ, intraepithelial or invasion of lamina propria

T1

Tumor invades the submucosa

T2

Tumor invades the muscularis propria

T3

Tumor invades through the muscularis propria into the subserosa, or into the nonperitonealized pericolic or perirectal tissues

T4

Tumor directly invades other organs or structures and/or perforates visceral peritoneum. We followed the International Union Against Cancer (UICC) classification

Supplementary material

535_2010_279_MOESM1_ESM.tif (2.9 mb)
Supplementary Fig. 1 Partial epitope mapping of anti-DK mAbs. One microgram of purified GST fusion proteins GST-hDK-β(22–167), GST-hDK-β(168–346) and GST-hDK-β(347–410) of DK-β were subjected to SDS-PAGE, stained with Coomassie Brilliant Blue (CBB) and immunoblotted with anti-DK mAb #45. The monoclonal Ab #45 exclusively recognized GST hDK-β(22–167), which indicates that its epitope is located between amino acids 22 and 167 of human DK-β; Ab #45 did not react with isoforms hDK-α, -ε or -δ (TIFF 2.91 mb)
535_2010_279_MOESM2_ESM.tif (1.5 mb)
Supplementary Fig. 2 ROC curve for DK(Tis-T1). Receiver-operator characteristic (ROC) curves for DK in separating normal healthy control and early-stage (Tis-T1) colorectal cancer patients. The area under curve was 67.0% (TIFF 1.48 mb)
535_2010_279_MOESM3_ESM.tif (122 kb)
Supplementary Table 1 Relationship between clinicopathological features and serum DK in colorectal cancer patients. The χ2 test was performed to determine correlations among the various parameters, and Fisher's exact test was used as appropriate. Results are expressed as means ± SD. Differences were considered significant when the two-tailed P was <0.05. Well, well-differentiated adenocarcinoma; pap, papillary adenocarcinoma; tubl, well-differentiated type; Mod, moderately differentiated adenocarcinoma; tub2,moderately differentiated type; Por, poorly differentiated adenocarcinoma (TIFF 121 kb)
535_2010_279_MOESM4_ESM.tif (503 kb)
Supplementary Table 2 The serum concentartion of DK, CAE, CA19-9 and S-p53in adenoma. The only one case showed high level of DK in adenoma, DK test did not include high false-positives in our data. We speculated that the expression of DK in adenoma did not affect serum concentration (TIFF 503 kb)
535_2010_279_MOESM5_ESM.doc (62 kb)
Supplementary Material and Methods (DOC 62 kb)

Copyright information

© Springer 2010

Authors and Affiliations

  • Tomoyuki Tagi
    • 1
  • Takeshi Matsui
    • 2
  • Shojiro Kikuchi
    • 1
    • 3
  • Sachi Hoshi
    • 4
  • Toshiya Ochiai
    • 1
  • Yukihito Kokuba
    • 1
  • Yoko Kinoshita-Ida
    • 5
  • Fumie Kisumi-Hayashi
    • 5
  • Koji Morimoto
    • 6
    • 7
  • Toshio Imai
    • 5
  • Issei Imoto
    • 8
    • 9
  • Johji Inazawa
    • 9
  • Eigo Otsuji
    • 1
  1. 1.Department of SurgeryKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Medical Top Track (MTT) Program, Medical Research InstituteTokyo Medical and Dental UniversityTokyoJapan
  3. 3.Department of SurgeryHyogo College of MedicineNishinomiyaJapan
  4. 4.Eisai Tsukuba Research Laboratories, Eisai Co., Ltd.TsukubaJapan
  5. 5.KAN Research Institute, Inc.KobeJapan
  6. 6.Department of Breast and Endocrine Surgery, Graduate School of MedicineOsaka UniversitySuitaJapan
  7. 7.Department of BiochemistryGraduate School of Medicine Osaka UniversitySuitaJapan
  8. 8.Department of Human Genetics and Public Health, Graduate School of Medical ScienceThe University of TokushimaTokushimaJapan
  9. 9.Department of Molecular Cytogenetics, Medical Research InstituteTokyo Medical and Dental UniversityTokyoJapan