Secretome analysis of oral keratinocytes chronically exposed to shisha

被引:4
作者
Patil, Shankargouda [1 ,2 ]
Babu, Niraj [3 ,4 ]
Subbannayya, Tejaswini [3 ]
Mohan, Sonali, V [3 ]
Sathe, Gajanan [3 ]
Solanki, Hitendra S. [3 ]
Rajagopalan, Pavithra [3 ]
Patel, Krishna [3 ]
Advani, Jayshree [3 ]
Bhandi, Shilpa [5 ]
Sidransky, David [6 ]
Chatterjee, Aditi [3 ,4 ]
Gowda, Harsha [3 ,4 ]
Ferrari, Marco [1 ,7 ]
机构
[1] Univ Siena, Dept Med Biotechnol, Sch Dent Med, Siena, Italy
[2] Jazan Univ, Dept Maxillofacial Surg & Diagnost Sci, Div Oral Pathol, Coll Dent, Jazan, Saudi Arabia
[3] Int Technol Pk, Inst Bioinformat, Bangalore 560066, Karnataka, India
[4] Manipal Acad Higher Educ, Manipal, Karnataka, India
[5] Jazan Univ, Dept Restorat Dent Sci, Div Operat Dent, Coll Dent, Jazan, Saudi Arabia
[6] Johns Hopkins Univ, Sch Med, Dept Otolaryngol Head & Neck Surg, Baltimore, MD 21205 USA
[7] Univ Leeds, Dept Restorat Dent, Sch Dent, Leeds, W Yorkshire, England
关键词
OKF6/TERT1; secretory biomarkers; waterpipe; TMT; oral cancer; proteomics; SQUAMOUS-CELL CARCINOMA; POTENTIAL BIOMARKERS; MATRIX METALLOPROTEINASES; DECREASED EXPRESSION; CIGARETTE-SMOKE; RISK-FACTORS; CANCER; PROTEIN; HEAD; IDENTIFICATION;
D O I
10.3233/CBM-182099
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
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.
引用
收藏
页码:29 / 41
页数:13
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