Download PDF
CC BY-NC-ND 4.0 · Ann Natl Acad Med Sci 2023; 59(03): 152-157
DOI: 10.1055/s-0043-1772214
Original Article

Cystatin A Down-regulation in Head and Neck Squamous Cell Carcinoma Cell Lines Decreases Cancer Hallmark Signatures

Geeta S. Boora
1   Department of Biochemistry, Post Graduate Institute of Medical Education and Research, Chandigarh, India
,
Anshika Chauhan
1   Department of Biochemistry, Post Graduate Institute of Medical Education and Research, Chandigarh, India
,
Amanjeet Bal
2   Department of Histopathology, Post Graduate Institute of Medical Education and Research, Chandigarh, India
,
Roshan K. Verma
3   Department of ENT, Post Graduate Institute of Medical Education and Research, Chandigarh, India
,
Arnab Pal 
1   Department of Biochemistry, Post Graduate Institute of Medical Education and Research, Chandigarh, India
› Author Affiliations Funding This work was supported by PGIMER Special Research Grant for funding the study and Council for Scientific and Industrial Research (CSIR), New Delhi, for providing the fellowship to AC.
› Further Information
   Permissions and Reprints

Abstract

Background Cystatin A (CSTA), an endogenous inhibitor of lysosomal cysteine protease, is expressed primarily in epithelial tissues. The expression of CSTA was found to be dysregulated in various cancers and associated with cancer pathogenesis, but its role is reported to be contradictory. Our previous preliminary study found CSTA to be upregulated in the saliva and tissues of patients with head and neck squamous cell carcinoma (HNSCC). In this current study, we have explored the role of CSTA in the pathophysiology of HNSCC.

Methods First, we confirmed the upregulation of CSTA in CAL 27 (p = 0.0242) and FaDu (p = 0.0014), two HNSCC cell lines, compared to the normal gingival epithelium. CSTA was then stably knocked down in CAL 27 and FaDu using the lentiviral short hairpin RNA pLKO vector transduction to study the effects of CSTA knockdown on various cancer hallmarks such as cell proliferation ability, invasion, migration, colony formation, and chemotherapy-induced apoptosis.

Results CSTA knockdown significantly decreased cell viability, cell migration, transwell invasion, and colony formation in both cell lines. CSTA downregulation also enhanced cisplatin-induced apoptosis.

Conclusion Overall, this study suggests the protumorigenic role of CSTA in HNSCC.

Keywords

cystatin A - HNSCC - shRNA pLKO vector transduction - cancer hallmarks

Ethics Approval Statement

This study was performed after getting the approval of the Institute Ethics Committee, PGIMER, Chandigarh.


Data Availability Statement

Detailed data and protocols shall be made available by the corresponding author on reasoned request.


Authors' Contribution

G.S.B., A.C., and A.P. conceived and planned the experiments. G.S.B. carried out the experiments. G.S.B. wrote the manuscript with input from all authors. A.B. analyzed the IHC and ICC slides. G.S.B., A.C., A.B, R.K.V., and A.P. contributed to the analysis and interpretation of the results. A.C. and A.P. edited the manuscript


Supplementary Material



Publication History

Article published online:
21 August 2023

© 2023. National Academy of Medical Sciences (India). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 

  • References

  • 1 Sung H, Ferlay J, Siegel RL. et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71 (03) 209-249
    CrossrefPubMedGoogle Scholar
  • 2 Polanska H, Raudenska M, Gumulec J. et al. Clinical significance of head and neck squamous cell cancer biomarkers. Oral Oncol 2014; 50 (03) 168-177
    CrossrefPubMedGoogle Scholar
  • 3 Barrett AJ. The cystatins: a diverse superfamily of cysteine peptidase inhibitors. Biomed Biochim Acta 1986; 45 (11-12): 1363-1374
    PubMedGoogle Scholar
  • 4 Brzin J, Kopitar M, Turk V, Machleidt W. Protein inhibitors of cysteine proteinases. I. Isolation and characterization of stefin, a cytosolic protein inhibitor of cysteine proteinases from human polymorphonuclear granulocytes. Hoppe Seylers Z Physiol Chem 1983; 364 (11) 1475-1480
    CrossrefPubMedGoogle Scholar
  • 5 Sinha AA, Quast BJ, Wilson MJ. et al. Ratio of cathepsin B to stefin A identifies heterogeneity within Gleason histologic scores for human prostate cancer. Prostate 2001; 48 (04) 274-284
    CrossrefPubMedGoogle Scholar
  • 6 Chang KP, Wu CC, Chen HC. et al. Identification of candidate nasopharyngeal carcinoma serum biomarkers by cancer cell secretome and tissue transcriptome analysis: potential usage of cystatin A for predicting nodal stage and poor prognosis. Proteomics 2010; 10 (14) 2644-2660
    CrossrefPubMedGoogle Scholar
  • 7 Kos J, Krasovec M, Cimerman N, Nielsen HJ, Christensen IJ, Brünner N. Cysteine proteinase inhibitors stefin A, stefin B, and cystatin C in sera from patients with colorectal cancer: relation to prognosis. Clin Cancer Res 2000; 6 (02) 505-511
    PubMedGoogle Scholar
  • 8 Roesch-Ely M, Nees M, Karsai S. et al. Proteomic analysis reveals successive aberrations in protein expression from healthy mucosa to invasive head and neck cancer. Oncogene 2007; 26 (01) 54-64
    CrossrefPubMedGoogle Scholar
  • 9 Budihna M, Strojan P, Smid L. et al. Prognostic value of cathepsins B, H, L, D and their endogenous inhibitors stefins A and B in head and neck carcinoma. Biol Chem Hoppe Seyler 1996; 377 (06) 385-390 F
    CrossrefPubMedGoogle Scholar
  • 10 Luo A, Kong J, Hu G. et al. Discovery of Ca2+-relevant and differentiation-associated genes downregulated in esophageal squamous cell carcinoma using cDNA microarray. Oncogene 2004; 23 (06) 1291-1299
    CrossrefPubMedGoogle Scholar
  • 11 Shiba D, Terayama M, Yamada K. et al. Clinicopathological significance of cystatin A expression in progression of esophageal squamous cell carcinoma. Medicine (Baltimore) 2018; 97 (15) e0357
    CrossrefPubMedGoogle Scholar
  • 12 Yang Z, Liao J, Carter-Cooper BA, Lapidus RG, Cullen KJ, Dan H. Regulation of cisplatin-resistant head and neck squamous cell carcinoma by the SRC/ETS-1 signaling pathway. BMC Cancer 2019; 19 (01) 485
    CrossrefPubMedGoogle Scholar
  • 13 Strojnik T, Zajc I, Bervar A. et al. Cathepsin B and its inhibitor stefin A in brain tumors. Pflugers Arch 2000; 439 (3, Suppl): R122-R123
    CrossrefPubMedGoogle Scholar
  • 14 Ebert E, Werle B, Jülke B. et al. Expression of cysteine protease inhibitors stefin A, stefin B, and cystatin C in human lung tumor tissue. Adv Exp Med Biol 1997; 421: 259-265
    CrossrefPubMedGoogle Scholar
  • 15 Kuopio T, Kankaanranta A, Jalava P. et al. Cysteine proteinase inhibitor cystatin A in breast cancer. Cancer Res 1998; 58 (03) 432-436
    PubMedGoogle Scholar
  • 16 Leto G, Tumminello FM, Pizzolanti G, Montalto G, Soresi M, Gebbia N. Lysosomal cathepsins B and L and Stefin A blood levels in patients with hepatocellular carcinoma and/or liver cirrhosis: potential clinical implications. Oncology 1997; 54 (01) 79-83
    CrossrefPubMedGoogle Scholar
  • 17 Butler MW, Fukui T, Salit J. et al. Modulation of cystatin A expression in human airway epithelium related to genotype, smoking, COPD, and lung cancer. Cancer Res 2011; 71 (07) 2572-2581
    CrossrefPubMedGoogle Scholar
  • 18 Jones B, Roberts PJ, Faubion WA, Kominami E, Gores GJ. Cystatin A expression reduces bile salt-induced apoptosis in a rat hepatoma cell line. Am J Physiol 1998; 275 (04) G723-G730
    Google Scholar