Identification a novel clinical biomarker in early diagnosis of human non-small cell lung cancer
- 70 Downloads
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.
KeywordsNon-small cell lung cancer Biomarker Dimethylation labeling Lectin AANL Serum paraoxonase/arylesterase 1 Alpha-1-antichymotrypsin
non-small cell lung cancer
squamous cell carcinoma
tumour node metastasis
agrocybe aegerita lectin 2 (high affinity to GlcNAc)
- PNGase F
peptide N-glycosidase F
horse radish peroxidase
serum paraoxonase/arylesterase 1
receiver operator characteristic
area under curve
enzyme-linked immunosorbent assay
bovine serum albumin
differentially expressed protein
healthy and benign
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).
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.
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.
- 14.Molina, R., Filella, X.: Aug, eacute, M., J., Fuentes, R., Bover, I., Rifa, J., Moreno, V., canals, E., vi, ntilde, olas, N., Marquez, a., Barreiro, E., Borras, J., Viladiu, P.: tumor markers (CEA, CA 125, CYFRA 21-1, SCC and NSE) in patients with non-small cell lung Cancer as an aid in histological diagnosis and prognosis. Tumor Biol. 24(4), 209–218 (2003)CrossRefGoogle Scholar
- 17.Shen, S., Zhang, R., Guo, Y., Loehrer, E., Wei, Y., Zhu, Y., Yuan, Q., Moran, S., Fleischer, T., Bjaanaes, M.M., Karlsson, A., Planck, M., Staaf, J., Helland, A., Esteller, M., Su, L., Chen, F., Christiani, D.C.: A multi-omic study reveals BTG2 as a reliable prognostic marker for early-stage non-small cell lung cancer. Mol. Oncol. 12, 913–924 (2018)CrossRefGoogle Scholar
- 21.Ahn, J.M., Sung, H.J., Yoon, Y.H., Kim, B.G., Yang, W.S., Lee, C., Park, H.M., Kim, B.J., Lee, S.Y., An, H.J., Cho, J.Y.: Integrated glycoproteomics demonstrates fucosylated serum paraoxonase 1 alterations in small cell lung cancer. Molecular & Cellular Proteomics : MCP. 13(1), 30–48 (2014)CrossRefGoogle Scholar
- 22.Liu, Y., Wang, C., Wang, R., Wu, Y., Zhang, L., Liu, B.F., Cheng, L., Liu, X.: Isomer-specific profiling of N-glycans derived from human serum for potential biomarker discovery in pancreatic cancer. J. Proteome. 181, 160-169 (2018)Google Scholar
- 24.Drake, P.M., Schilling, B., Niles, R.K., Prakobphol, A., Li, B., Jung, K., Cho, W., Braten, M., Inerowicz, H.D., Williams, K., Albertolle, M., Held, J.M., Iacovides, D., Sorensen, D.J., Griffith, O.L., Johansen, E., Zawadzka, A.M., Cusack, M.P., Allen, S., Gormley, M., Hall, S.C., Witkowska, H.E., Gray, J.W., Regnier, F., Gibson, B.W., Fisher, S.J.: Lectin chromatography/mass spectrometry discovery workflow identifies putative biomarkers of aggressive breast cancers. J. Proteome Res. 11(4), 2508–2520 (2012)CrossRefGoogle Scholar
- 25.Satomaa, T., Heiskanen, A., Leonardsson, I., Angstrom, J., Olonen, A., Blomqvist, M., Salovuori, N., Haglund, C., Teneberg, S., Natunen, J., Carpen, O., Saarinen, J.: Analysis of the human cancer glycome identifies a novel group of tumor-associated N-acetylglucosamine glycan antigens. Cancer Res. 69(14), 5811–5819 (2009)CrossRefGoogle Scholar
- 27.Jin, Y., Wang, J., Ye, X., Su, Y., Yu, G., Yang, Q., Liu, W., Yu, W., Cai, J., Chen, X., Liang, Y., Chen, Y., Wong, B.H., Fu, X., Sun, H.: Identification of GlcNAcylated alpha-1-antichymotrypsin as an early biomarker in human non-small-cell lung cancer by quantitative proteomic analysis with two lectins. Br. J. Cancer. 114(5), 532–544 (2016)CrossRefGoogle Scholar
- 30.Li, L., Song, J., Kalt, W., Forney, C., Tsao, R., Pinto, D., Chisholm, K., Campbell, L., Fillmore, S., Li, X.: Quantitative proteomic investigation employing stable isotope labeling by peptide dimethylation on proteins of strawberry fruit at different ripening stages. J. Proteome. 94, 219–239 (2013)CrossRefGoogle Scholar
- 32.Zeng, G.Q., Zhang, P.F., Deng, X., Yu, F.L., Li, C., Xu, Y., Yi, H., Li, M.Y., Hu, R., Zuo, J.H., Li, X.H., Wan, X.X., Qu, J.Q., He, Q.Y., Li, J.H., Ye, X., Chen, Y., Li, J.Y., Xiao, Z.Q.: Identification of candidate biomarkers for early detection of human lung squamous cell cancer by quantitative proteomics. Mol. Cell. Proteomics : MCP. 11(6), M111 013946 (2012)Google Scholar
- 36.Elkiran, E.T., Mar, N., Aygen, B., Gursu, F., Karaoglu, A., Koca, S.: Serum paraoxonase and arylesterase activities in patients with lung cancer in a Turkish population. BMC Cancer. 7(48), (2007)Google Scholar
- 39.Atay, A.E., Kaplan, M.A., Evliyaoglu, O., Ekin, N., Isikdogan, A.: The predictive role of Paraoxonase 1 (PON1) activity on survival in patients with metastatic and nonmetastatic gastric cancer. Clin. Ter. 165(1), e1–e5 (2014)Google Scholar
- 40.Keskin, M., Dolar, E., Dirican, M., Kiyici, M., Yilmaz, Y., Gurel, S., Nak, S.G., Erdinc, S., Gulten, M.: Baseline and salt-stimulated paraoxonase and arylesterase activities in patients with chronic liver disease: relation to disease severity. Intern. Med. J. 39(4), 243–248 (2009)CrossRefGoogle Scholar
- 41.Huang, C., Wang, Y., Liu, S., Ding, G., Liu, W., Zhou, J., Kuang, M., Ji, Y., Kondo, T., Fan, J.: Quantitative proteomic analysis identified paraoxonase 1 as a novel serum biomarker for microvascular invasion in hepatocellular carcinoma. J. Proteome Res. 12(4), 1838–1846 (2013)CrossRefGoogle Scholar
- 42.Sun, C., Chen, P., Chen, Q., Sun, L., Kang, X., Qin, X., Liu, Y.: Serum paraoxonase 1 heteroplasmon, a fucosylated, and sialylated glycoprotein in distinguishing early hepatocellular carcinoma from liver cirrhosis patients. Acta Biochim. Biophys. Sin. Shanghai. 44(9), 765–773 (2012)CrossRefGoogle Scholar