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Serum exosome-derived biomarkers for the early detection of oral squamous cell carcinoma

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Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Blood exosomes help regulate communication between tumour cells, moderating their behaviour. We sought to determine the protein content in serum exosomes (SEs), to characterise SEs, and to discover novel clinical biomarkers of oral squamous cell carcinoma (OSCC). Differentially expressed proteins (DEPs) of OSCC were identified using proteomics and then analysed using bioinformatics, before validation using ELISA, IHC, and RT-PCR. The influence of SEs on oral cancer cells was detected using CCK-8 and migration assays. Twelve DEPs were found in SEs from OSCC. Four proteins were targeted for further verification. New biomarkers exhibiting high sensitivity and specificity in diagnosing OSCC comprised C-reactive protein (CRP), von willebrand factor (VWF), and leucine-rich alpha-2-glycoprotein (LRG). Combined biomarkers outperformed any single protein. We also demonstrated that tumour-derived exosomes promoted tumour cell migration, but not proliferation and apoptosis. Our study indicates that CRP, VWF, and LRG are potential clinically relevant OSCC biomarkers. OSCC-related SEs may help promote migration of oral cells.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

OSCC:

Oral squamous cell carcinomas

DEPs:

Differential expressed proteins

CRP:

C-reactive protein

VWF:

Von Willebrand factor

LRG1:

Leucine-rich alpha-2-glycoprotein

OC:

Oral cancer

HCs:

Healthy controls

MS:

Mass spectrometry

GO:

Gene ontology

PPI network:

Protein interaction network

STRING:

Search tool for the retrieval of interacting genes

KEGG:

Kyoto encyclopedia of genes and genomes pathways

LNM:

Lymph node metastasis

NLNM:

No lymph node metastasis

C1QA:

Complement C1q subcomponent subunit A

GEPIA:

Gene expression profiling interactive analysis

ROCs:

Receiver operation characteristic curves

AUC:

The area under the curve

CCK-8:

Cell Counting Kit-8

PI:

Propidium iodide

BP:

Biological processes

CC:

Cellular component

MF:

Molecular functions

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Acknowledgements

The authors would like to thank all the reviewers who participated in the review and MJEditor (www.mjeditor.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This work is supported by grants from Guangxi medical and health appropriate technology research and development project (Grant Nos.: S201538), Guangxi Clinical Research Center for Craniofacial Deformity (Grant Nos.: GKAD17129004) and National Natural Science Foundation of China (81670970, 81870748).

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Authors and Affiliations

  1. Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment, Guangxi Health Commission Key Laboratory of Prevention and Treatment for Oral Infectious Diseases, Guangxi Clinical Research Center for Craniofacial Deformity, College of Stomatology, Guangxi Medical University, Nanning, 530021, People’s Republic of China

    Hejia Guo, Weidong Jiang, Suhua Huang, Xuanping Huang & Cuiping Li

  2. Department of Oral and Maxillofacial Surgery, College & Hospital of Stomatology, Guangxi Medical University, Nanning, 530021, People’s Republic of China

    Hejia Guo, Weidong Jiang & Xuanping Huang

  3. Medical Scientific Research Center, College of Stomatology, Guangxi Medical University, Nanning, 530021, People’s Republic of China

    Cuiping Li

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  1. Hejia Guo
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Contributions

HJG, CPL-collected serum samples; HJG, WDJ, SHH-conducted mass spectrometry analysis, verification tests and statistical analysis; HJG, CPL, XPH-wrote the manuscript.

Corresponding authors

Correspondence to Xuanping Huang or Cuiping Li.

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All authors declare that they have no conflicts of interests.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee of Guangxi Medical University (the committee’s reference number: 2018-082). Informed consent was obtained from all individual participants included in the study.

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Supplementary Information

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11010_2021_4254_MOESM1_ESM.tif

Supplementary file1: Mass spectra of differentially-expressed proteins. The identified peptide fragments of four biomarkers including the b-ion and y-ion series. A CRP; B LRG; C VWF; D C1QA (TIF 731 kb)

11010_2021_4254_MOESM2_ESM.tif

Supplementary file2: RT-PCR validation of MS/MS data. A1-A2/B1-B2/C1-C2/D1-D2 CRP/LRG/VWF/C1QA mRNA expression levels in tissues assayed by RT-PCR. LNM: oral squamous cell carcinoma with lymph node metastasis; NLNM: oral squamous cell carcinoma with no lymph node metastasis; CT: cancerous tissues; CAT: corresponding adjacent tissues. Data were shown as mean ± SD. (nonparametric test, p < 0 .05 was considered statistically significant) (TIF 25514 kb)

11010_2021_4254_MOESM3_ESM.tif

Supplementary file3: Clinical interrelation from RT-PCR results of tissue samples. A1-A7, B1-B7, C1-C7, D1-D7 represented the expressions of CRP, LRG, VWF, C1QA in gender, nationality, stage, differentiation degree, smoking, drinking, and the grades of age brackets. WDSCC: Well differentiated squamous-cell carcinoma; LDSCC: Low differentiated squamous-cell carcinoma. Data were shown as mean ± SD. (Chi-square test, McNemar and Fisher’s exact tests, p < 0.05 was considered statistically significant) (TIF 25515 kb)

11010_2021_4254_MOESM4_ESM.tif

Supplementary file4: The results from GEPIA analysis. Gene expression profile, stage plots, and survival plots showed the connection between CRP, LRG, VWF, and C1QA and head and neck squamous cell carcinoma. A CRP; B LRG; C VWF; D C1QA (TIF 61956 kb)

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Guo, H., Jiang, W., Huang, S. et al. Serum exosome-derived biomarkers for the early detection of oral squamous cell carcinoma. Mol Cell Biochem 476, 4435–4447 (2021). https://doi.org/10.1007/s11010-021-04254-7

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