Proteome profiling to identify peroxiredoxin 1 interacting protein partners in nicotine-associated oral leukoplakia.

Abstract

OBJECTIVE:

Tobacco smoking is one of the main risk factors for oral squamous cell carcinoma (OSCC) and can induce generation of reactive oxygen species (ROS). In our previous studies, we demonstrated that nicotine, the major ingredient in tobacco, can upregulate an important antioxidant enzyme Peroxiredoxin 1 (Prx1), in oral leukoplakia (OLK), an oral precancerous lesion. The underlying regulatory mechanisms, however, remain unclear. This study aims to identify regulatory mechanisms of nicotine and identify Prx1 interacting proteins in nicotine-associated OLK.

DESIGN:

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with bioinformatics analysis was conducted to profile Prx1 binding proteins in human dysplastic oral keratinocyte (DOK) cells. Candidate interaction proteins were further verified using Co-immunoprecipitation (Co-IP), Western blot or Duolink assay in 4-nitro-quinoline-1-oxide (4NQO)-induced OLK in mice and human OLK tissues.

RESULTS:

We identified Thioredoxin (Trx), Nucleolar GTP-binding protein 1 (GTPBP4), GTP-binding protein Di-Ras2 (DIRAS2) and apoptosis signal-regulating kinase 1 (ASK1) as key Prx1 interacting proteins regulated by nicotine. Our data showed that nicotine upregulated Trx, GTPBP4, DIRAS2, and downregulated ASK1 in 4NQO-induced OLK in mice, at least in part dependent on Prx1. The modulations of Trx, GTPBP4, DIRAS2 and ASK1 by nicotine were also found in OLK smokers compared to OLK non-smokers. The in-situ interaction of Trx, GTPBP4, DIRAS2 and ASK1 with Prx1 were validated in human OLK tissues.

CONCLUSION:

Nicotine may promote OLK development via regulating Prx1 binding proteins Trx, GTPBP4, DIRAS2 and ASK1. The results of this study will help to develop therapeutic approaches for OLK in humans targeting Prx1 interacting protein network.

• Authors: Moci Qi, Lingyu Li, Yunping Lu, Hui Chen, Min Zhang, Min Wang, Lihua Ge, Jing Yang, Ni Shi, Tong Chen, Xiaofei Tang
• Journal: Archives of oral biology (Arch Oral Biol) Vol. 108 Pg. 104537 (Dec 2019) ISSN: 1879-1506 [Electronic] England
• PMID: 31525533 (Publication Type: Journal Article)
• Copyright: Published by Elsevier Ltd.

Chemical References

• DIRAS3 protein, human
• Homeodomain Proteins
• Nuclear Proteins
• PRRX1 protein, human
• Proteome
• Nicotine
• Peroxiredoxins
• MAP Kinase Kinase Kinase 5
• MAP3K5 protein, human
• GTP-Binding Proteins
• GTPBP4 protein, human
• rho GTP-Binding Proteins

Topics

• Animals
• Carcinoma, Squamous Cell (metabolism)
• Chromatography, Liquid
• GTP-Binding Proteins
• Homeodomain Proteins (metabolism)
• Humans
• Leukoplakia, Oral
• MAP Kinase Kinase Kinase 5 (metabolism)
• Mice
• Mouth Neoplasms (metabolism)
• Nicotine (pharmacology)
• Nuclear Proteins
• Peroxiredoxins
• Proteome
• Smoking (adverse effects)
• Tandem Mass Spectrometry
• rho GTP-Binding Proteins (metabolism)