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High Co-expression of Large Tenascin C Splice Variants in Stromal Tissue and Annexin A2 in Cancer Cell Membranes is Associated with Poor Prognosis in Pancreatic Cancer

  • Kei Hagiwara
  • Norifumi HarimotoEmail author
  • Takehiko Yokobori
  • Ryo Muranushi
  • Kouki Hoshino
  • Dorgormaa Gantumur
  • Takahiro Yamanaka
  • Norihiro Ishii
  • Mariko Tsukagoshi
  • Takamichi Igarashi
  • Hiroshi Tanaka
  • Akira Watanabe
  • Norio Kubo
  • Kenichiro Araki
  • Yasuo Hosouchi
  • Ken Shirabe
Translational Research and Biomarkers

Abstract

Background

Pancreatic cancer tissue contains abundant stromal components, including extracellular matrix proteins such as tenascin C (TNC), which exists as large (TNC-L) and non-large splice variants. Here, we examined human pancreatic cancer specimens for the expression of total TNC (TNC-ALL) and TNC-L in the stroma and annexin A2 (ANXA2), a cell surface receptor for TNC, and evaluated their significance as prognostic markers for pancreatic cancer.

Methods

Expression of ANXA2, TNC-ALL, and TNC-L was examined in 106 pancreatic cancer tissues from patients who underwent curative resection and who had not received prior therapy or surgery. Protein expression was measured by immunohistochemistry and scored on a semi-quantitative scale. The relationships between protein expression, clinicopathological factors, and prognosis were evaluated by Cox proportional hazards analysis.

Results

TNC-ALL and TNC-L were detected mainly in the stroma, whereas ANXA2 was predominantly expressed in cancer cell membranes. TNC-ALL was also expressed in non-tumor pancreatic tissue. High levels of stromal TNC-L and membranous ANXA2, but not stromal TNC-ALL, were independently associated with cancer progression and poor prognosis. Moreover, high co-expression of stromal TNC-L and membranous ANXA2 was a superior indicator of poor prognosis compared with detection of TNC-ALL, TNC-L, or ANXA2 alone.

Conclusions

Our data suggest that co-expression of stromal TNC-L and membranous ANXA2 is a poor prognostic marker compared with detection of TNC-L or ANXA2 alone for pancreatic cancer patients. Additionally, targeting of crosstalk between stromal TNC and cancer cell ANXA2 could be a promising therapeutic strategy to overcome refractory pancreatic cancer.

Notes

Acknowledgment

The authors thank Anne M. O’Rourke, PhD, and H. Nikki March, PhD, from Edanz Group (www.edanzediting.com/ac) for editing a draft of this manuscript.

Disclosure

The authors have no conflicts of interest directly relevant to the content of this article.

Supplementary material

10434_2019_7708_MOESM1_ESM.tif (425 kb)
Supplementary Fig. 1 Immunohistochemical staining of stromal TNC-ALL and TNC-L in pancreatic cancer tissue. Left: Positive staining of TNC-ALL. Right: Absence of TNC-L staining. Original magnification ×200 (TIFF 425 kb)
10434_2019_7708_MOESM2_ESM.tif (173 kb)
Supplementary Fig. 2 Disease-free survival curves of pancreatic cancer patients stratified by expression of stromal TNC-ALL and TNC-L and tumor cell ANXA2. (a–f) Kaplan–Meier curves showing disease-free survival of patients stratified by expression or co-expression of proteins. a High vs. low stromal TNC-ALL (P = 0.0192). b High vs. low stromal TNC-L (P = 0.001). c Stromal TNC-NL vs. TNC-L expression (P = 0.0030) and TNC-NL vs. TNC-negative (P = 0.4540). d High vs. low membranous expression of ANXA2 (P = 0.0020). e High stromal TNC-ALL and high membranous ANXA2 co-expression vs. low stromal TNC-ALL and low membranous ANXA2 co-expression (P < 0.001). f High stromal TNC-L and high membranous ANXA2 co-expression vs. low stromal TNC-L and low membranous ANXA2 co-expression (P < 0.001) (TIFF 172 kb)
10434_2019_7708_MOESM3_ESM.tif (170 kb)
Supplementary Fig. 3 Overall survival and disease-free survival curves of pancreatic cancer patients at each clinical stage (stage I–II and III–IV) stratified by expression of stromal TNC-ALL and TNC-L and cancer cell ANXA2. a Overall survival (stage IA–IIB): high vs. low stromal TNC-ALL (P = 0.017). b Overall survival (stage IA–IIB): high vs. low stromal TNC-L (P ≤ 0.001). c Overall survival (stage IA–IIB): high vs. low membranous expression of ANXA2 (P = 0.0020). d Overall survival (stage III–IV): high vs. low stromal TNC-ALL (P = 0.609). e Overall survival (stage II–IV): high vs. low stromal TNC-L (P = 0.798). f Overall survival (stage III–IV): high vs. low membranous expression of ANXA2 (P = 0.039) (TIFF 170 kb)
10434_2019_7708_MOESM4_ESM.tif (134 kb)
Supplementary Fig. 4 Overall survival and disease-free survival curves of pancreatic cancer patients stratified by cytoplasmic expression of TNC-ALL and TNC-L. a Overall survival: high vs. low cytoplasm TNC-ALL (P = 0.669). b Overall survival: high vs. low cytoplasmic TNC-L (P = 0.586). c Disease-free survival: high vs. low cytoplasmic TNC-ALL (P = 0.787). d Disease-free survival: high vs. low cytoplasmic TNC-ALL (P = 0.586) (TIFF 134 kb)
10434_2019_7708_MOESM5_ESM.tif (92 kb)
Supplementary Fig. 5 Overall survival curves of pancreatic cancer patients with high or low ANXA2 expression stratified by expression of TNC-ALL and TNC-L in the stroma. a Overall survival (ANXA2 high): high vs. low stromal TNC-ALL (P = 0.060). b Overall survival (ANXA2 high): high vs. low stromal TNC-L (P < 0.001). c Overall survival (ANXA2 low): high vs. low stromal TNC-ALL (P = 0.351). d Overall survival (ANXA2 low): high vs. low stromal TNC-L (P = 0.144) (TIFF 92 kb)
10434_2019_7708_MOESM6_ESM.tif (89 kb)
Supplementary material 6 (TIFF 88 kb)
10434_2019_7708_MOESM7_ESM.tif (155 kb)
Supplementary material 7 (TIFF 154 kb)
10434_2019_7708_MOESM8_ESM.tif (173 kb)
Supplementary material 8 (TIFF 172 kb)

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Copyright information

© Society of Surgical Oncology 2019

Authors and Affiliations

  • Kei Hagiwara
    • 1
  • Norifumi Harimoto
    • 1
    Email author
  • Takehiko Yokobori
    • 2
    • 3
  • Ryo Muranushi
    • 1
  • Kouki Hoshino
    • 1
  • Dorgormaa Gantumur
    • 1
  • Takahiro Yamanaka
    • 1
  • Norihiro Ishii
    • 1
  • Mariko Tsukagoshi
    • 1
    • 2
  • Takamichi Igarashi
    • 1
  • Hiroshi Tanaka
    • 1
  • Akira Watanabe
    • 1
  • Norio Kubo
    • 1
  • Kenichiro Araki
    • 1
  • Yasuo Hosouchi
    • 4
  • Ken Shirabe
    • 1
  1. 1.Department of Hepatobiliary and Pancreatic SurgeryGunma University, Graduate School of MedicineMaebashiJapan
  2. 2.Department of Innovative Cancer ImmunotherapyGunma UniversityMaebashiJapan
  3. 3.Gunma University Initiative for Advanced Research (GIAR)MaebashiJapan
  4. 4.Department of Surgery and Laparoscopic SurgeryGunma Prefecture Saiseikai Maebashi HospitalMaebashiJapan

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