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Journal of Cell Communication and Signaling

, Volume 13, Issue 3, pp 281–289 | Cite as

Chronic shisha exposure alters phosphoproteome of oral keratinocytes

  • Shankargouda Patil
  • Pavithra Rajagopalan
  • Krishna Patel
  • Tejaswini Subbannayya
  • Niraj Babu
  • Sonali V. Mohan
  • Jayshree Advani
  • Gajanan Sathe
  • Shilpa Bhandi
  • Hitendra S. Solanki
  • David Sidransky
  • Aditi Chatterjee
  • Harsha GowdaEmail author
  • Marco FerrariEmail author
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Abstract

Shisha smoking has been epidemiologically linked to oral cancer. However, few studies have investigated the pathobiology of shisha-induced cellular transformation. We studied the effects of chronic shisha exposure (8 months) in an in vitro model using immortalized, non-neoplastic oral keratinocytes (OKF6/TERT1). Quantitative proteomic and phosphoproteomic analyses were performed on OKF6/TERT1 cells treated with shisha extract for a period of 8 months. Pathway analysis was carried out to identify significantly enriched biological processes in shisha-treated cells. Chronic shisha exposure resulted in increased cell scattering phenomenon in OKF6/TERT1 cells. Data analysis revealed differential phosphorylation of 164 peptides (fold change ≥1.5, p ≤ 0.0.5) corresponding to 136 proteins. Proteins associated with mTORC1 and EIF4F complexes involved in initiating protein translation were seen to be enriched upon shisha treatment. Network analysis also highlighted downregulation of proteins involved in Type I interferon signaling in shisha-treated cells. Quantitative phosphoproteomic approach elucidated global perturbations to the molecular milieu of oral keratinocytes upon shisha exposure. Further studies are needed to validate putative targets in oral cancer patients with shisha smoking history.

Keywords

Waterpipe Hookah Narghile Orbitrap fusion High-throughput 

Notes

Acknowledgements

We thank the Department of Biotechnology (DBT), Government of India for research support to the Institute of Bioinformatics. IOB is supported by DBT Program Support on Neuroproteomics and infrastructure for proteomic data analysis (BT/01/COE/08/05). Krishna Patel and Niraj Babu are recipients of Senior Research Fellowships from the Council for Scientific and Industrial Research (CSIR), Government of India. We thank Dr. Anita Mahadevan of National Institute of Mental Health and Neurosciences (NIMHANS) for providing use of microscope facility.

Author’s contributions

AC, HG and SP participated in study conception and study design. PR, TS, NB and HS were involved in cell culture and performed all assays and experiments. SVM and GS carried out fractionation and mass spectrometric analysis of samples. PR and KP prepared the manuscript and manuscript figures. PR, KP and JA were involved in data analyses and interpretation. AC, HG, SP, DS, SB and MF edited, critically read and revised the manuscript. All the authors have read and approved the final manuscript.

Compliance with ethical standards

Approved by ethics committee/institute

Not applicable.

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

12079_2019_528_MOESM1_ESM.docx (15 kb)
ESM 1 (DOCX 15 kb)

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Copyright information

© The International CCN Society 2019

Authors and Affiliations

  • Shankargouda Patil
    • 1
    • 2
  • Pavithra Rajagopalan
    • 3
  • Krishna Patel
    • 3
  • Tejaswini Subbannayya
    • 3
  • Niraj Babu
    • 3
    • 4
  • Sonali V. Mohan
    • 3
    • 4
    • 5
  • Jayshree Advani
    • 3
  • Gajanan Sathe
    • 3
  • Shilpa Bhandi
    • 6
  • Hitendra S. Solanki
    • 3
  • David Sidransky
    • 7
  • Aditi Chatterjee
    • 3
    • 4
  • Harsha Gowda
    • 3
    • 4
    Email author
  • Marco Ferrari
    • 2
    • 8
    Email author
  1. 1.Department of Maxillofacial Surgery and Diagnostic Sciences, Division of Oral Pathology, College of DentistryJazan UniversityJazanSaudi Arabia
  2. 2.Department of Medical Biotechnologies, School of Dental MedicineUniversity of SienaSienaItaly
  3. 3.Institute of Bioinformatics, International Technology ParkBangaloreIndia
  4. 4.Manipal Academy of Higher Education (MAHE)ManipalIndia
  5. 5.Department of Genetics and Computational Biology, QIMR Berghofer Medical Research InstituteBrisbaneAustralia
  6. 6.Department of Restorative Dental Sciences, Division of Operative Dentistry, College of DentistryJazan UniversityJazanSaudi Arabia
  7. 7.Department of Otolaryngology-Head and Neck SurgeryJohns Hopkins University School of MedicineBaltimoreUSA
  8. 8.Department of Restorative Dentistry, School of DentistryUniversity of LeedsLeedsUK

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