In-gel digestion coupled with mass spectrometry (GeLC-MS/MS)-based salivary proteomic profiling of canine oral tumors

Sekkarin Ploypetch,Narumon Phaonakrop,Sittiruk Roytrakul,Chanin Kalpravidh,Suthathip Kittisenachai,Sirinun Pisamai,Anudep Rungsipipat,Kantinan Leetanasaksakul,Gunnaporn Suriyaphol

doi:10.1186/s12917-020-02550-w

Abstract

BackgroundVarious types of oral tumors, either benign or malignant, are commonly found in dogs. Since saliva directly contacts the tumors and saliva collection is non-invasive, easily accessible and cost effective, salivary biomarkers are practical to be used for the diagnosis and/or prognosis of these diseases. However, there is limited knowledge of protein expression in saliva for canine oral tumors. The present study aimed to investigate novel biomarkers from the salivary proteome of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), and periodontitis and healthy controls (CP), using an in-gel digestion coupled with mass spectrometry (GeLC-MS/MS). The relationships between protein candidates and chemotherapy drugs were explored and the expression of potential biomarkers in saliva and tissues was verified by western blot analysis.ResultsFor saliva samples, increased expression of protein tyrosine phosphatase non-receptor type 5 (PTPN5) was shown in all tumor groups compared with the CP group. Marked expression of PTPN5 was also observed in LOM and OSCC compared with that in BN and EOM. In addition, tumor protein p53 (p53), which appeared in the PTPN5–drug interactions, was exhibited to be expressed in all tumor groups compared with that in the CP group. For tissue samples, increased expression of p53 was shown in LOM compared with the control group.ConclusionPTPN5 and p53 were proposed to be potential salivary biomarkers of canine oral tumors.

Highlights

Various types of oral tumors, either benign or malignant, are commonly found in dogs
The present study aimed to search for novel suitable biomarkers in saliva of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), periodontitis (P) and healthy controls (C) (CP group), using in-gel digestion coupled with mass spectrometry (GeLC-Mass spectrometry (MS)/MS)
The distribution of the individual and overlapped proteins in early-stage oral melanoma (EOM), LOM, OSCC, BN and controls and periodontitis (CP) groups was illustrated by a Venn diagram (Fig. 1)

Summary

Introduction

Various types of oral tumors, either benign or malignant, are commonly found in dogs. The present study aimed to investigate novel biomarkers from the salivary proteome of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), and periodontitis and healthy controls (CP), using an in-gel digestion coupled with mass spectrometry (GeLC-MS/MS). The relationships between protein candidates and chemotherapy drugs were explored and the expression of potential biomarkers in saliva and tissues was verified by western blot analysis. The present study aimed to search for novel suitable biomarkers in saliva of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), periodontitis (P) and healthy controls (C) (CP group), using in-gel digestion coupled with mass spectrometry (GeLC-MS/MS). The candidate protein expressions in saliva and tissues were affirmed by western blot analysis

Methods

Results

Discussion

Conclusion