Cellular Oncology

, Volume 41, Issue 2, pp 185–200 | Cite as

Role of β-catenin in cisplatin resistance, relapse and prognosis of head and neck squamous cell carcinoma

  • Souvick Roy
  • Madhabananda Kar
  • Shomereeta Roy
  • Arka Saha
  • Swatishree Padhi
  • Birendranath Banerjee
Original Paper

Abstract

Background

Head and neck squamous cell carcinoma (HNSCC) is one of the most common types of cancer in India with high incidence and rapid recurrence rates. Here, we aimed to investigate the role of β-catenin, a developmental pathway gene, in HNSCC therapy resistance, DNA damage response, recurrence and prognosis.

Methods

In total 80 HNSCC samples were included. Western blot, immunohistochemistry and qRT-PCR analyses were performed to assess β-catenin expression in the cut margin and tumor areas of each sample. Kaplan-Meier analyses were performed to correlate β-catenin expression with the survival and prognosis of HNSCC patients. In addition, chemo-resistance, DNA damage response and DNA repair capacities were evaluated in HNSCC-derived cell lines through LiCl-mediated up-regulation and siRNA-mediated silencing of β-catenin expression.

Results

We observed β-catenin up-regulation in cut margin areas of recurrent patients compared to their corresponding tumor regions, which subsequently could be associated with poor prognosis. In addition, we found that LiCl-mediated up-regulation of β-catenin in HNSCC-derived cells led to cisplatin resistance, evasion of apoptosis, enhanced DNA repair and enhanced migration. The effects of β-catenin silencing correlated with its putative role in chemo-resistance and DNA damage response.

Conclusion

From our results we conclude that β-catenin may contribute to HNSCC therapy resistance and disease relapse. As such, β-catenin may be explored as a therapeutic target along with conventional therapeutics.

Keywords

Head and neck squamous cell carcinoma (HNSCC) β catenin Cisplatin-resistance Disease relapse DNA damage repair and response Prognosis 

Notes

Acknowledgements

This work was supported by a grant from the Department of Atomic Energy (DAE), Board of Research for Nuclear Sciences (BRNS), Government of India, Grant Number 2013/35/45/BRNS and by the MSSB (Molecular Stress and Stem Cell Biology) group.

Compliance with ethical standard

Ethical approval

This study was approved by the institutional ethics committee of the School of Biotechnology and Kalinga Institute of Medical Sciences (KIMS), KIIT University, and was conducted according to the Helsinki declaration. The human sample collection was carried out strictly according to the institutional ethical board guidelines.

Conflict of interest

The authors declare no conflict of interest.

Informed consent

Informed consent was obtained from all subjects or their nominees prior to participation in the study.

Supplementary material

13402_2017_365_Fig8_ESM.gif (64 kb)
Supplementary Fig. 1

Silencing of β-catenin in HCT-116 cells. (a) mRNA level expression of β-catenin in HCT-116 cells after transfected with scrambled siRNA and siRNA- β-catenin. (b) Representative western blot results and graphical representation of β-catenin expression in scrambled siRNA and siRNA-β-catenin transfected HCT-116 cells. (GIF 64 kb)

13402_2017_365_MOESM1_ESM.tif (571 kb)
High Resolution Image (TIFF 571 kb)
13402_2017_365_Fig9_ESM.gif (675 kb)
Supplementary Fig. 2

β-catenin is plays an important role in chemoresistance in HCT-116 cells. (a) Cell viability assay of parental, scrambled-siRNA transfected and siRNA-β-catenin transfected HCT-116 cells after treatment with different concentrations of cisplatin. (b,c) Determination of colony forming capacity and cell survival of parental, scrambled-siRNA and siRNA-β-catenin transfected HCT-116 cells after treatment with different concentrations of cisplatin by clonogenic assay. (d) Graphical representation of percentage apoptotic cells (Sub G0) in parental, scrambled-siRNA and siRNA-β-catenin transfected HCT-116 cells after cisplatin treatment. (e) Representative image of wound in parental, scrambled-siRNA and siRNA-β-catenin transfected cells at 0 h and 24 h after cisplatin treatment. (f) Graphical representation of percentage wound closure after 24 h of cisplatin treatment in parental, scrambled-siRNA and siRNA-β-catenin transfected HCT-116 cells. (GIF 674 kb)

13402_2017_365_MOESM2_ESM.tif (7.5 mb)
High Resolution Image (TIFF 7638 kb)
13402_2017_365_Fig10_ESM.gif (120 kb)
Supplementary Fig. 3

Silencing of β-catenin in UPCI-SCC-131 HNSCC cell line and its effect on cell cycle progression. (a) UPCI-SCC-131 cells were transfected with scrambled siRNA and siRNA- β-catenin for 48 h. mRNA expression of β-catenin in scrambled siRNA and siRNA- β-catenin transfected UPCI-SCC-131 cells . (b) Representative western blot results of β-catenin expression in scrambled siRNA and siRNA-β-catenin transfected UPCI-SCC-131 cells. (c) Graphical representation of relative protein expression of β-catenin in scrambled-siRNA and siRNA-β-catenin transfected UPCI-SCC-131 cells. (d) Representative histograms of cell cycle analysis in scrambled-siRNA transfected cells and siRNA-β-catenin transfected cells. (e) Graphical representation of percentage distribution of cells in scrambled-siRNA transfected cells and siRNA-β-catenin transfected UPCI-SCC-131 cells. (GIF 120 kb)

13402_2017_365_MOESM3_ESM.tif (1.2 mb)
High Resolution Image (TIFF 1270 kb)
13402_2017_365_MOESM4_ESM.pdf (167 kb)
Supplementary Table 1 Sequences of the primers used in this study. (PDF 166 kb)
13402_2017_365_MOESM5_ESM.pdf (16 kb)
Supplementary Table 2 Clinico-pathological characteristics of 80 HNSCC patients. (PDF 15 kb)
13402_2017_365_MOESM6_ESM.pdf (9 kb)
Supplementary Table 3 Association of co-morbidity factors with recurrence in HNSCC patients. (PDF 8 kb)

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Copyright information

© International Society for Cellular Oncology 2017

Authors and Affiliations

  • Souvick Roy
    • 1
  • Madhabananda Kar
    • 2
  • Shomereeta Roy
    • 1
  • Arka Saha
    • 1
  • Swatishree Padhi
    • 1
  • Birendranath Banerjee
    • 1
  1. 1.Molecular Stress and Stem Cell Biology Group, School of BiotechnologyKIIT UniversityBhubaneswarIndia
  2. 2.Department of Surgical OncologyAll India Institute of Medical Sciences (AIIMS)BhubaneswarIndia

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