Comparative sera proteomics analysis of differentially expressed proteins in oral squamous cell carcinoma

Yin-Ling Wong; Anand Ramanathan; Kar Mun Yuen; Wan Mahadzir Wan Mustafa; Mannil Thomas Abraham; Keng Kiong Tay; Zainal Ariff Abdul Rahman; Yeng Chen

doi:10.7717/peerj.11548

Comparative sera proteomics analysis of differentially expressed proteins in oral squamous cell carcinoma

PeerJ. 2021 Jun 10:9:e11548. doi: 10.7717/peerj.11548. eCollection 2021.

Authors

Yin-Ling Wong¹, Anand Ramanathan^{2

3}, Kar Mun Yuen⁴, Wan Mahadzir Wan Mustafa⁵, Mannil Thomas Abraham⁴, Keng Kiong Tay⁴, Zainal Ariff Abdul Rahman^{2

3}, Yeng Chen^{1

3}

Affiliations

¹ Department of Oral & Craniofacial Sciences, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia.
² Department of Oral & Maxillofacial Clinical Sciences, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia.
³ Oral Cancer Research & Coordinating Centre, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia.
⁴ Oral Health Program, Ministry of Health, Federal Government Administrative Centre, Putrajaya, Malaysia.
⁵ Faculty of Dentistry, MAHSA University, Jenjarum, Selangor, Malaysia.

Abstract

Background: Oral squamous cell carcinoma (OSCC) has increased in incidence from 1990 to 2017, especially in South and Southeast Asia. It is often diagnosed at an advanced stage with a poor prognosis. Therefore, early detection of OSCC is essential to improve the prognosis of OSCC. This study aims to identify the differentially expressed serum proteins as potential biomarkers for oral squamous cell carcinoma (OSCC).

Methods: Comparative proteomics profiling of serum samples from OSCC patients, oral potentially malignant disorder (OPMD) patients, and healthy individuals were performed using two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry (MS) (n = 60) and bioinformatics analysis. The enzyme-linked immunosorbent assay (ELISA) (n = 120) and immunohistochemistry (IHC) (n = 70) were used to confirm our findings.

Results: The 2-DE analysis revealed that 20 differentially expressed proteins were detected in OPMD and OSCC (p < 0.05). Bioinformatics analysis indicated that the activation of classical complement, liver X receptor/retinoid X receptor (LXR/RXR) activation, and acute phase response signaling pathway are associated with the development and progression of OSCC. Most of the detected proteins are acute-phase proteins and were related to inflammation and immune responses, including apolipoprotein A-I (APOA1), complement C3 (C3), clusterin (CLU), and haptoglobin (HP). The expression levels of CLU and HP in ELISA are consistent with the findings from the 2-DE analysis, except for the mean serum level of HP in OPMD, whereby it was slightly higher than that in control. IHC results demonstrated that CLU and HP are significantly decreased in OSCC tissues.

Conclusion: Decreased expression of CLU and HP could serve as complementary biomarkers of OSCC. These proteins may assist in predicting the outcomes of OSCC patients. However, a larger cohort is needed for further investigation.

Keywords: Biomarker; Oral potentially malignant disorder; Oral squamous cell carcinoma; Proteomics.

Grants and funding

This work was supported by the University of Malaya Postgraduate Research Grant (PPP) PG326-2016A and University of Malaya (UM) High Impact Research (HIR) MoE Grants UM.C/625/1/HIR/MOE/DENT/09 and UM.C/625/1/HIR/MOHE/MED/16/5 from the Ministry of Education Malaysia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.