Integrated Proteomics Based on 2D Gel Electrophoresis and Mass Spectrometry with Validations: Identification of a Biomarker Compendium for Oral Submucous Fibrosis-An Indian Study.

Divyambika Catakapatri Venugopal,Soundharya Ravindran,Sathasivasubramanian Sankarapandian,Vidyarani Shyamsundar,Arvind Krishnamurthy,Vijayalakshmi Ramshankar,Ananthi Sivagnanam,Yasasve Madhavan

doi:10.3390/jpm12020208

Abstract

Oral Submucous Fibrosis (OSMF) is a chronic debilitating disease more frequently found in the South East Asian population. This disease poses a public health priority, as it is grouped under oral potentially malignant disorders, with malignant transformation rates of around 7 to 13%. Hence, early identification of high-risk OSMF patients is of the utmost importance to prevent malignant transformation. Proteomic expression profiling is a promising method for identifying differentially expressed proteins for disease prognosis and risk stratification in OSMF. In this study, overexpressed proteins in OSMF, OSMF transformed into oral squamous cell carcinoma (OSCC) and normal tissues were evaluated by proteomic analysis using two-dimensional electrophoresis (2DE) and mass spectrometry, which revealed 23 upregulated proteins. Validation was done using immunohistochemistry for three secretory proteins, namely 14-3-3ε (n = 130), carbonic anhydrase 1 (CA 1) (n = 125) and heat shock protein 70 (HSP 70) (n = 117), which showed significant overexpression in OSMF, OSCC compared to normal. The present study is the first of its kind in India to the best of our knowledge, assessing the altered expression of proteins in OSMF and OSMF which has undergone malignant transformation, obtaining a better knowledge of the molecular pathways involved in the disease progression. The current study shows that the biomarkers studied can be potentially useful for risk stratification of OSMF to OSCC serving as novel targets for therapeutic intervention. Clinical validation of the targets can further pave way for precision medicine to improve the quality of life in OSMF patients.

Highlights

Our study shows comprehensive differentially expressed profiles of Oral Submucous Fibrosis (OSMF) and OSMF along with oral squamous cell carcinoma (OSCC) in the same sample compared with the normal samples
A comparative proteomic study of pooled tissue samples was taken from patients with OSMF, OSMF with histologically proven OSCC and was compared to pooled normal samples (Figure 1)
The current study has shown that the discovery of protein biomarkers based on proteomic analysis can help in the assessment of disease prognosis and aid in the development of targeted therapy, as described in existing literature [27,28]

Summary

Introduction

Oral submucous fibrosis is a chronic, potentially malignant disorder of the oral mucosa, which is more widespread in the Indian subcontinent [1,2]. The main etiological determinant is the areca nut, which stimulates fibroblast proliferation and collagen synthesis and reduces collagen breakdown. It consists of alkaloids, namely arecoline, arecaidine, guvacine and guvacoline, which undergo nitrosation to form nitrosamines, which further alkylates with DNA, leading to malignant transformation on prolonged exposure. Listed as a Group I carcinogen by the International Agency for Research on Cancer, the areca nut is a common component of betel quid, which is predominantly used in Southeast Asia [3]

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Discussion

Conclusion