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Identification a novel clinical biomarker in early diagnosis of human non-small cell lung cancer

  • Yanxia Jin
  • Yajun Yang
  • Yanting Su
  • Xiangdong Ye
  • Wei Liu
  • Qing Yang
  • Jie Wang
  • Xiangning Fu
  • Yongsheng GongEmail author
  • Hui SunEmail author
Original Article
  • 72 Downloads

Abstract

Non-small cell lung cancer (NSCLC) is a malignant tumor with high morbidity and mortality. The clinical biomarkers currently used for the early diagnosis of lung cancer have poor sensitivity and specificity. Therefore, it is urgent to identify sensitive biomarkers for the early detection of NSCLC to improve the patient survival of patients. In our previously study, we identified glycoprotein alpha-1-antichymotrypsin (AACT) as an early biomarker of NSCLC. In this study, serum glycopeptides were enriched using the high-GlcNAc-specific binding lectin, AANL/AAL2, for further quantitative proteomics analysis using LC-MS/MS. A total of 55 differentially expressed proteins were identified by using demethylation labelling proteomics. Serum paraoxonase/arylesterase 1 (PON1) was selected for validation by western blotting and lectin-ELISA in samples from 120 enrolled patients. Our data showed that AANL-enriched PON1 has better diagnostic performance than total PON1 in early NSCLC, since it differed between early Stage I tumor samples and tumor-free samples (healthy and benign). Combining AANL-enriched PON1 with carcinoembryonic antigen (CEA) significantly improved the diagnostic specificity of CEA. Moreover, combined AANL-enriched PON1 and AANL-enriched AACT was significantly different between early NSCLC samples and tumor-free samples with an AUC of 0.940, 94.4% sensitivity, and 90.2% specificity. Our findings suggest that combined AANL-enriched PON1 and AANL-enriched AACT is a potential clinical biomarker for the early diagnosis of NSCLC.

Keywords

Non-small cell lung cancer Biomarker Dimethylation labeling Lectin AANL Serum paraoxonase/arylesterase 1 Alpha-1-antichymotrypsin 

Abbreviations

NSCLC

non-small cell lung cancer

AD

adenocarcinoma

SCC

squamous cell carcinoma

TNM

tumour node metastasis

GlcNAc

β-N-acetyl-D-glucosamine

AALNL/AAL2

agrocybe aegerita lectin 2 (high affinity to GlcNAc)

TOF

time-of-flight

GO

gene ontology

PNGase F

peptide N-glycosidase F

HRP

horse radish peroxidase

PON1

serum paraoxonase/arylesterase 1

AACT

alpha-1-antichymotrypsin

ROC

receiver operator characteristic

AUC

area under curve

ELISA

enzyme-linked immunosorbent assay

CEA

carcinoembryonic antigen

BSA

bovine serum albumin

MS

mass spectrum

DEPs

differentially expressed protein

HB

healthy and benign

Notes

Acknowledgements

We would like to thank Professor Xiangdong Fu (University of California, San Diego, USA) for his helpful suggestions on the study. We thank Pengfei Cheng in Wuhan University Hospital and Wangjie in Tongji Medical Hospital for collection of serum samples. We thank Haoyu Li, Feilong Yu and Xi Chen for protein labeling and analysis of MS data. This work was supported by the Natural Science Foundation of China (NSFC) program [grant numbers No. 81670531, 31370849, 31800676], Jiangsu Natural Science Foundation program (grant number No. BK20141176), the Chinese 111 project [grant number No. B06018], and Hong Kong Scholars Program (grant number No. XJ2018060).

Author’s contribution

YX. Jin designed the experiments, performed most experiments, analyzed the data, and wrote the manuscript. YJ. Yang performed the western blot experiments with assistance from YX. Jin. YT. Su and XD. Ye contributed to enrichment with lectin and lectin-elisa. W. Liu contribute to provide the PNGase F enzyme. Q. Yang contribute to provide the lectin AAL2. J. Wang and XN. Fu contributed to clinical patient information management and provided all clinical patient serum samples. H. Sun and YS. Gong provided overall project supervision and revised the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors have stated that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Supplementary material

10719_2018_9853_MOESM1_ESM.pdf (213 kb)
ESM 1 (PDF 213 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yanxia Jin
    • 1
    • 2
  • Yajun Yang
    • 1
  • Yanting Su
    • 1
  • Xiangdong Ye
    • 1
  • Wei Liu
    • 1
  • Qing Yang
    • 1
  • Jie Wang
    • 3
  • Xiangning Fu
    • 3
  • Yongsheng Gong
    • 4
    Email author
  • Hui Sun
    • 1
    • 5
    Email author
  1. 1.Hubei Key Laboratory of Cell Homeostasis, College of Life SciencesWuhan UniversityWuhanPeople’s Republic of China
  2. 2.Hubei Key Laboratory of Edible Wild Plants Conservation and UtilizationHubei Normal UniversityHuangshiPeople’s Republic of China
  3. 3.Tongji Medical HospitalHuazhong University of Science and TechnologyWuhanPeople’s Republic of China
  4. 4.Suzhou Municipal Hospitalthe Affiliated Suzhou Hospital of Nanjing Medical UniversitySuzhouPeople’s Republic of China
  5. 5.Hubei Province Key Laboratory of Allergy and ImmunologyWuhan UniversityWuhanPeople’s Republic of China

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