Journal of Cancer Research and Clinical Oncology

, Volume 143, Issue 2, pp 243–254 | Cite as

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

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.

Keywords

Gli1 CSC Esophageal squamous cell carcinoma Biomarker 

Notes

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).

Compliance with ethical standards

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.

Supplementary material

432_2016_2273_MOESM1_ESM.tif (12.4 mb)
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 (1.2 mb)
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)
432_2016_2273_MOESM3_ESM.doc (126 kb)
Supplementary material 3 (DOC 126 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Key Laboratory of Natural Resources of the Changbai Mountain and Functional Molecules, Ministry of EducationYanbian UniversityYanjiChina
  2. 2.Institute for Regenerative MedicineYanbian University College of MedicineYanjiChina
  3. 3.Department of OncologyAffiliated Hospital of Yanbian UniversityYanjiChina
  4. 4.Department of Pathology, Samsung Medical CenterSungkyunkwan University College of MedicineSeoulKorea

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