Skip to main content
SpringerLink
Log in

Gli1, a potential regulator of esophageal cancer stem cell, is identified as an independent adverse prognostic factor in esophageal squamous cell carcinoma

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

The hedgehog (Hh) pathway is involved in cancer stem cell (CSC) maintenance in various tumors. Glioma-associated oncogene homolog 1 (Gli1) is a key mediator of the Hh pathway; however, its expression and clinical significance in esophageal squamous cell carcinoma (ESCC) have not been reported. In this study, we aimed to reveal clinical significance of Gli1 expression in ESCC and further investigate the potential of Gli1 as a CSC regulator of ESCC by comparing its expression with expressions of other stemness genes in ESCC.

Methods

We assessed the expressions of Gli1, Sox9, CD44, Sox2, LSD1, and Oct4 in 127 patients’ tissue specimens of ESCC using immunohistochemistry and in ESCC cell lines using Western blotting. The relationship of Gli1 expression with clinic–pathologic parameters as well as cell-cycle-regulating genes was investigated. We also investigated the biological pathways that are activated in Gli1-high ESCC using The Cancer Genome Atlas (TCGA) data.

Results

Gli1 expression was observed in 28.3 % of ESCC, and its expression was correlated with the expression of stemness genes, Sox9 (P = 0.003) and CD44 (P = 0.012). And Gli1, CD44, and Sox9 were highly expressed in more poorly differentiated ESCC cell lines such as TE8 and TE1 cells. Notably, Gli1 expression was positively associated with distant metastasis (P = 0.011), increased microvessel density (MVD) (P = 0.002), and expression of cell cycle regulators such as p21, cyclin D1, cyclin E1, and NF-κB (P < 0.05). Sox9 and CD44 expressions in ESCC were also significantly associated with unfavorable clinic–pathologic parameters such as increased MVD, advanced tumor (pT) stage, and higher TNM stage. Moreover, all three potential CSC markers such as Gli1, Sox9, and CD44 were strongly linked to worse clinical outcome and independent poor prognostic factors in overall survival and disease-free survival in ESCC. Gene set enrichment analysis revealed that the Gli1-high-expressing ESCC patients’ group was strongly enriched for gene expression signature of Hh signaling pathway, epithelial–mesenchymal transition, and cancer stem cell.

Conclusions

Targeting Gli1, a potential diagnostic marker of ESCC stem cells, will have a profound therapeutic and prognostic value.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abbas T, Dutta A (2009) p21 in cancer: intricate networks and multiple activities. Nat Rev Cancer 9(6):400–414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Boopalan T, Arumugam A, Parada J et al (2015) Receptor activator for nuclear factor-κB ligand signaling promotes progesterone-mediated estrogen-induced mammary carcinogenesis. Cancer Sci 106:25–33

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bora-Singhal N, Perumal D, Nguyen J et al (2015) Gli1-mediated regulation of Sox2 facilitates self-renewal of stem-like cells and confers resistance to EGFR inhibitors in non-small cell lung cancer. Neoplasia 17(7):538–551

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Clement V, Sanchez P, de Tribolet N et al (2007) HEDGEHOG-GLI1 signaling regulates human glioma growth, cancer stem cell self-renewal, and tumorigenicity. Curr Biol 17:165–172

    Article  CAS  PubMed  Google Scholar 

  • Colavito SA, Zou MR, Yan Q et al (2014) Significance of glioma-associated oncogene homolog 1 (GLI1) expression in claudin-low breast cancer and crosstalk with the nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) pathway. Breast Cancer Res 16:444–462

    Article  PubMed  PubMed Central  Google Scholar 

  • Cui D, Chen X, Yin J et al (2012) Aberrant activation of Hedgehog/Gli1 pathway on angiogenesis in gliomas. Neurol India 60:589–596

    Article  PubMed  Google Scholar 

  • de Carcer G, Perez DCI, Malumbres M (2007) Targeting cell cycle kinases for cancer therapy. Curr Med Chem 14(9):969–985

    Article  PubMed  Google Scholar 

  • Deng W, Vanderbilt DB, Lin CC et al (2015) SOX9 inhibits β-TrCP-mediated protein degradation to promote nuclear GLI1 expression and cancer stem cell properties. J Cell Sci 128(6):1123–1138

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Du L, Wang H, He L et al (2008) CD44 is of functional importance for colorectal cancer stem cells. Clin Cancer Res 14:6751–6760

    Article  CAS  PubMed  Google Scholar 

  • Gotoda T, Matsumura Y, Kondo H et al (2000) Expression of CD44 variants and prognosis in oesophageal squamous cell carcinoma. Gut 46:14–19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guo JF, Gao J, Li Z, Gong Y et al (2013) Adenovirus vector-mediated Gli1 siRNA induces growth inhibition and apoptosis in human pancreatic cancer with Smo-dependent or Smo-independent Hh pathway activation in vitro and in vivo. Cancer Lett 339:185–194

    Article  CAS  PubMed  Google Scholar 

  • Hong Y, Chen W, Du X et al (2015) Upregulation of sex-determining region Y-box 9 (SOX9) promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma. Oncotarget 10:18632–18646

    Google Scholar 

  • Islam F, Gopalan V, Wahab R et al (2015) Cancer stem cells in oesophageal squamous cell carcinoma: identification, prognostic and treatment perspectives. Oncology 96:9–19

    Google Scholar 

  • Lange CA, Yee D (2011) Killing the second messenger: targeting loss of cell cycle control in endocrine-resistant breast cancer. Endocr Relat Cancer 18(4):19–24

    Article  Google Scholar 

  • Min S, Xiaoyan X, Fanghui P et al (2013) The glioma-associated oncogene homolog 1 promotes epithelial–mesenchymal transition in human esophageal squamous cell cancer by inhibiting E-cadherin via Snail. Cancer Gene Ther 20:379–385

    Article  CAS  PubMed  Google Scholar 

  • O’Toole SA, Machalek DA, Shearer RF et al (2011) Hedgehog overexpression is associated with stromal interactions and predicts for poor outcome in breast cancer. Cancer Res 71(11):4002–4014

    Article  PubMed  Google Scholar 

  • Qi YJ, Chao WX, Chiu JF (2012) An overview of esophageal squamous cell carcinoma proteomics. J Proteomics 75:3129–3137

    Article  CAS  PubMed  Google Scholar 

  • Senzer N, Barve M, Kuhn J et al (2012) Phase I trial of “bi-shRNAi(furin)/GMCSF DNA/autologous tumor cell” vaccine (FANG) in advanced cancer. Mol Ther 20:679–686

    Article  CAS  PubMed  Google Scholar 

  • Sheng W, Dong M, Zhou J et al (2014) The clinicopathological significance and relationship of Gli1, MDM2 and p53 expression in resectable pancreatic cancer. Histopathology 64:523–535

    Article  PubMed  Google Scholar 

  • Souzaki M, Kubo M, Kai M et al (2011) Hedgehog signaling pathway mediates the progression of noninvasive breast cancer to invasive breast cancer. Cancer Sci 102:373–381

    Article  CAS  PubMed  Google Scholar 

  • Sun L, Yu S (2011) Meta-analysis: non-steroidal anti-inflammatory drug use and the risk of esophageal squamous cell carcinoma. Dis Esophagus 24:544–549

    Article  PubMed  Google Scholar 

  • Wang K, Pan L, Che X et al (2010) Gli1 inhibition induces cell-cycle arrest and enhanced apoptosis in brain glioma cell lines. J Neurooncol 98(3):319–327

    Article  CAS  PubMed  Google Scholar 

  • Xu M, Gong A, Yang H et al (2015) Sonic hedgehog-glioma associated oncogene homolog 1 signaling enhances drug resistance in CD44(+)/Musashi-1(+) gastric cancer stem cells. Cancer Lett 369(1):124–133

    Article  CAS  PubMed  Google Scholar 

  • Yang ZT, Yeo SY, Yin YX et al (2016) Tenascin-C, a prognostic determinant of esophageal squamous cell carcinoma. PLoS One 11(1):1–17

    Google Scholar 

  • Zhao JS, Li WJ, Ge D et al (2011) Tumor initiating cells in esophageal squamous cell carcinomas express high levels of CD44. PLoS One 6:1–11

    Google Scholar 

Download references

Acknowledgments

This study was supported by Grants from National Natural Science Funds of China (81460390) and Science and Technology Development Plan of Jilin Province Fund of China. And this study was supported by a Grant of the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (HI14C2517).

Author information

Author notes

    Authors and Affiliations

    1. Key Laboratory of Natural Resources of the Changbai Mountain and Functional Molecules, Ministry of Education, Yanbian University, Yanji, 133002, China

      Zhaoting Yang, Weidong Ni & Yanhua Xuan

    2. Institute for Regenerative Medicine, Yanbian University College of Medicine, Yanji, 133002, China

      Zhaoting Yang, Weidong Ni & Yanhua Xuan

    3. Department of Oncology, Affiliated Hospital of Yanbian University, Yanji, China

      Yan Cui

    4. Department of Pathology, Samsung Medical Center, Sungkyunkwan University College of Medicine, Seoul, 110-745, Korea

      Seokhyung Kim

    Authors
    1. Zhaoting Yang
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Yan Cui
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Weidong Ni
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Seokhyung Kim
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Yanhua Xuan
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding authors

    Correspondence to Seokhyung Kim or Yanhua Xuan.

    Ethics declarations

    Conflict of interest

    We declare that we have no conflict of interest.

    Ethics statement

    This research complied with the Helsinki Declaration and was approved by the Human Ethics Committee and the Research Ethics Committee of Samsung Medical Center. All patients provided written informed consent according to institutional guidelines. Patients were informed that the resected specimens were stored by the hospital and potentially used for scientific research and that their privacy would be maintained. Follow-up survival data were collected retrospectively through medical record analyses.

    Additional information

    Zhaoting Yang and Yan Cui have contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    432_2016_2273_MOESM1_ESM.tif

    Supplementary Fig. 1 Immunohistochemical staining of ESCC with LSD1, Sox2 and Oct4. Kaplan–Meier analyses of overall and disease-free survival curves for LSD1 (a and d), Sox2 (b and e) and Oct4 (c and f) expression in ESCC patients. (TIFF 12649 kb)

    432_2016_2273_MOESM2_ESM.tif

    Supplementary Fig. 2 Western blot analysis of LSD1, Sox2, and Oct44 in ESCC cells line. β-Actin was used as a loading control. (TIFF 1219 kb)

    Supplementary material 3 (DOC 126 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Yang, Z., Cui, Y., Ni, W. et al. Gli1, a potential regulator of esophageal cancer stem cell, is identified as an independent adverse prognostic factor in esophageal squamous cell carcinoma. J Cancer Res Clin Oncol 143, 243–254 (2017). https://doi.org/10.1007/s00432-016-2273-6

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00432-016-2273-6

    Keywords

    Search

    Navigation