Secretome analysis of oral keratinocytes chronically exposed to shisha

Abstract

BACKGROUND: Shisha smoking has been associated with multiple diseases including oral cancer. However, a mechanistic study to investigate alteration of secreted proteins in oral cells due to shisha smoking is lacking. OBJECTIVES: Elucidation of differentially secreted proteins by immortalized human normal oral keratinocytes (OKF6/TERT1) upon chronic exposure to shisha. METHODS: OKF6/TERT1 was chronically treated with 0.5% shisha extract for 8 months. Conditioned media from shisha treated (OKF6/TERT1-Shisha) and untreated (OKF6/TERT1-Parental) cells were subjected to TMT-based quantitative proteomic analysis. Bioinformatics analysis of differentially secreted proteins was carried out using SignalP, SecretomeP and TMHMM. Immunoblot validation of selected proteins was carried out to confirm the proteomics results. RESULTS: Proteomic analysis of OKF6/TERT1-Parental and OKF6/TERT1-Shisha secretome resulted in the identification of 1,598 proteins, of which 218 proteins were found to be differentially secreted (≥ 1.5-fold; p-value ≤ 0.05) in shisha treated cells. Bioinformatics analysis using prediction tools showed secretory potential of differentially secreted proteins identified in OKF6/TERT1-Shisha. Western blotting validated the expression of AKR1C2, HSPH1 and MMP9 in OKF6/TERT1-Shisha secretome in agreement with proteomic data. CONCLUSION: This study serves as a useful resource to understand the effect of chronic shisha smoking on the milieu of secreted proteins of oral cells. In vivo studies are warranted to supplement our in vitro data to elucidate the role of these proteins as early diagnostic biomarkers for oral carcinogenesis among shisha smokers.

Cytotoxic and Genotoxic Effects of Waterpipe on Oral Health Status

“…After screen reading and excluding paper unrelated to our search, 38 remained. Reports from the same authors/co-authors or same study center were excluded [14][15][16][17][18][19] as such reviews, comments, letters, hypotheses, and expert opinions 3,[20][21][22][23][24][25][26] . Two exclusive microbiological studies were removed 27,28 , same to animal studies 29,30 .…”

“…reviews, comments, letters, hypotheses and expert opinions). 3 , 14 – 26 Additionally, two exclusively microbiological studies were removed, and two were considered as animal studies. 27 – 30 However, one study was found but could not be retrieved properly.…”

Signaling alterations in oral keratinocytes in response to shisha and crude tobacco extract

“…Recently, a series of multiomic studies carried out by our group provided substantial evidence to indicate that chronic exposure to WS leads to oncogenic transformation in normal human oral keratinocytes. [5][6][7][8] The main component used in these studies is 'shisha'-a processed tobacco variant blended with flavors and additives.…”