Skip to main content
SpringerLink
Log in

EGFR/cell membrane chromatography-online-high performance liquid chromatography/mass spectrometry method for screening EGFR antagonists from Radix Angelicae Pubescentis

  • Articles
  • Published:
Science China Chemistry Aims and scope Submit manuscript

Abstract

The intracellular kinase domains of the epidermal growth factor receptor (EGFR) in some tumor cells are significant targets for drug discovery. We have developed a new EGFR cell membrane chromatography (EGFR/CMC)-online-high performance liquid chromatography/mass spectrometry (HPLC/MS) method for screening anti-EGFR antagonists from medicinal herbs such as Radix Angelicae Pubescentis. In this study, the HEK293 EGFR cells with high expression of EGFR were used to prepare cell membrane stationary phase (CMSP) in the EGFR/CMC model. The retention fractions on the EGFR/CMC model were directly analyzed by combining a 10 port columns switcher with a HPLC/MS system online. As a result, osthole from Radix Angelicae Pubescentis was found to be the active component acting on EGFR like dasatinib as the control drug. There was a good relationship between their inhibiting effects on EGFR secretion and HEK293 EGFR cell growth in vitro. This new EGFR/CMConline-HPLC/MS method can be applied for screening anti-EGFR antagonists from TCMs, for instance, Radix Angelicae Pubescentis. It will be a useful method for drug discovery with natural medicinal herbs as a leading compound resource.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Sugawa N, Ekstrand AJ, James CD, Collins VP. Identical splicing of aberrant epidermal growth factor receptor transcripts from amplified rearranged genes in human glioblastomas. Proc Natl Acad Sci USA, 1990, 87: 8602–8606

    Article  CAS  Google Scholar 

  2. Richard J, Sainsbury C, Needham GK, Farndon JR, Malcolm AJ, Harris AL. Epidermal-growth-factor receptor status as predictor of early recurrence of and death from breast cancer. Lancet, 1987, 329: 1398–1402

    Article  Google Scholar 

  3. Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med, 2004, 350: 2129–2139

    Article  CAS  Google Scholar 

  4. Radinsky R, Risin S, Fan D, Dong ZY, Bielenberg D, Bucana CD, Fidler IJ. Level and function of epidermal growth factor receptor predict the metastatic potential of human colon carcinoma cells. Clin Cancer Res, 1995, 1: 19–31

    CAS  Google Scholar 

  5. Eales-Reynolds LJ, Laver H, Mojtahedi H. Evidence for the expression of the EGFReceptor on human monocytic cells. Cytokine, 2001, 5: 169–172

    Article  Google Scholar 

  6. Kris MK, Natale RB, Herbst RS, Lynch TJ, Prager D, Belani CP, Schiller JH, Kelly K, Spiridonidis H, Sandler A, Albain KS, Cella D, Wolf MK, Averbuch SD, Ochs JJ, Kay AC. Efficacy of gefitinib, an inhibitor of the epidermal growth factor receptor tyrosine kinase, in symptomatic patients with non-small cell lung cancer. J Am Med Assoc, 2003, 290: 2149–2158

    Article  CAS  Google Scholar 

  7. Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nature Medicine, 2000, 6: 389–395

    Article  CAS  Google Scholar 

  8. Stamos J, Sliwkowski MX, Eigenbrot C. Structure of the epidermal growth factor receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor. J Biol Chem, 2002, 227: 46265–46272

    Article  Google Scholar 

  9. Ciardiello F, Tortora G. A novel approach in the treatment of cancer: targeting the epidermal growth factor receptor. Clin Cancer Res, 2001, 7: 2958–2970

    CAS  Google Scholar 

  10. Yang XD, Jia XC, Corvalan JRF, Wang P, Davis CG. Development of ABX-EGF, a fully human anti-EGF receptor monoclonal antibody, for cancer therapy. Crit Rev Oncol Hematol, 2001, 38: 17–23

    Article  CAS  Google Scholar 

  11. Wakeling AE, Guy SP, Woodburn JR, Ashton SE, Curry BJ, Barker AJ, Gibson KH. ZD1839 (Iressa): An orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. Cancer Res, 2002, 62: 5749–5754

    CAS  Google Scholar 

  12. Ullrich A, Coussens L, Hayflick JS, Dull TJ, Gray A, Tam AW, Lee J, Yarden Y, Libermann TA, Schlessinger J, Downward J, Mayes ELV, Whittle N, Waterfield MD, Seeburg PH. Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells. Nature, 309: 418–425

  13. Webb SED, Roberts SK, Needham SR, Tynan CJ, Rolfe DJ, Winn MD, Clarke DT, Barraclough R, Martin-Fernandez ML. Single-molecule imaging and fluorescence lifetime imaging microscopy show different structures for high- and low-affinity epidermal growth factor receptors in A431 cells. Biophysical J, 2008, 94: 803

    Article  CAS  Google Scholar 

  14. Schreiber SL. Target-oriented and diversity-oriented organic synthesis in drug discovery. Science, 2000, 287: 1964–1969

    Article  CAS  Google Scholar 

  15. Drews J. Drug discovery: a historical perspective. Science, 2000, 287: 1960–1964

    Article  CAS  Google Scholar 

  16. Kingsmore SF, Lindquist IE, Mudge J, Gessler DD, Beavis WD. Genome-wide association studies: progress and potential for drug discovery and development. Nat Rev Drug Discov, 2008, 7: 221–230

    Article  CAS  Google Scholar 

  17. Araujo RP, Liotta LA, Petricoin EF. Proteins, drug targets and the mechanisms they control: the simple truth about complex networks. Nat Rev Drug Discov, 2007, 6: 871–880

    Article  CAS  Google Scholar 

  18. He LC, Geng XD. Determination of ligustilide in rat blood and tissues by capillary gas chromatography/mass spectrometry. New Prog Biomed Chromatogr, 1996, 3, 8–9

    Google Scholar 

  19. He LC, Yang GD, Geng XD. Enzymatic activity and chromatographic characteristics of the cell membrane immobilized on silica surface. Chin Sci Bull, 1999, 44: 826–831

    Article  CAS  Google Scholar 

  20. He LC, Wang SC, Geng XD. Coating and fusing cell membranes onto a silica surface and their chromatographic characteristics. Chromatographia, 2001, 54: 71–76

    Article  CAS  Google Scholar 

  21. Hou XF, Zhou MZ, Jiang Q, Wang SC, He LC. A vascular smooth muscle/cell membrane chromatography-offline-gas chromatography/mass spectrometry method for recognition, separation and identification of active components from traditional Chinese medicines. J Chromatogr A, 2009, 1216(42): 7081–7087

    Article  CAS  Google Scholar 

  22. Yuan BX, Hou J, Yang GD, Zhao LM, He LC. Comparison of Determination of drug-muscarinic receptor affinity by cell-membrane chromatography and by radioligand-binding assay with the cerebrum membrane of the rat. Chromatographia, 2005, 61: 381–384

    Article  CAS  Google Scholar 

  23. Wang Y, Yuan BX, Deng XL, He LC, Zhang YY, Han QD. The preparation of HEK293 α1A or HEK293 α1B cell membrane stationary phase and the chromatographic affinity study of ligands of α1 adrenoceptor. Anal Biochem, 2005, 339: 198–205

    Article  CAS  Google Scholar 

  24. Wang Y, Yuan BX, Deng XL, He LC, Wang SC, Zhang YY, Han QD. Comparison of alpha1-adrenergic receptor cell-membrane stationary phases prepared from expressed cell line and from rabbit hepatocytes. Anal Bioanal Chem, 2006, 386: 2003–2011

    Article  CAS  Google Scholar 

  25. Zeng AG, Yuan BX, Wang CH, Yang GD, He LC. Frontal analysis of cell-membrane chromatography for determination of drug-alpha(1D) adrenergic receptor affinity. J Chromatogr B, 2009, 877(20–21): 1833–1837

    Article  CAS  Google Scholar 

  26. Zeng AG, Yuan BX, Zhu F, Zhao LM, He LC. Cell membrane chromatography correlated with functional assay for ligand-beta-adrenergic receptor affinities. Chromatographia, 2009, 69(11–12): 1373–1377

    Article  CAS  Google Scholar 

  27. Wang CH, He LC, Wang N, Yang GD, He LC. Screening anti-inflammatory components from Chinese traditional medicines using a peritoneal macrophage/cell membrane chromatography-offline-GC/MS method. J Chromatogr B, 2009, 877(27): 3019–3024

    Article  CAS  Google Scholar 

  28. Wang L, Ren J, Sun M, Wang SC. A combined cell membrane chromatography and online HPLC/MS method for screening compounds from Radix Caulophylli acting on the human α1A-adrenoceptor. J Pharm Biomed Anal, 2010, 51: 1302–1306

    Google Scholar 

  29. Wang SC, Wen BY, Wang L, Liu JT, He LC. Fluorenone alkaloid from Caulophyllum robustum Maxim with anti-myocardial ischemia activity. Arch Pharmacal Res, 2009, 32: 521–526

    Article  CAS  Google Scholar 

  30. Li CQ, He LC. Establishment of the model of white blood cell membrane chromatography and screening of antagonizing TLR4 receptor component from Atractylodes macrocephala Koidz. Sci China Ser C-Bio, 2006, 49(2): 182–189

    Article  CAS  Google Scholar 

  31. Li YP, He LC. Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application. Chin Sci Bull, 2007, 52: 922–928

    Article  CAS  Google Scholar 

  32. Li CQ, He LC, Dong HY, Jin JQ. Screening for the anti-inflammatory activity of fractions and compounds from Atractylodes macrocephala koidz. J Ethnopharmacol, 2007, 114: 212–217

    Article  CAS  Google Scholar 

  33. Liang MJ, He LC, Yang GD. Screening, analysis and in vitro vasodilatation of effective components from Ligusticum Chuanxiong. Life Sci, 2005, 78: 128–133

    Article  CAS  Google Scholar 

  34. He LC, Luo WJ, Li X, Xu XM, Wang SC, Ge XW, Li C. A wild-type EGFR expression of recombinant HEK293 cells. Chinese Patent, 200910022565.9

  35. Situ ZQ, Wu JZ. Cell Culture. Xi’an World Publishing Corporation. Xi’an, 2004, 241–250

    Google Scholar 

  36. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 1983, 65: 55–63

    Article  CAS  Google Scholar 

  37. Shimamura M, Hazato T, Ashino H, Yamamoto Y, Iwasaki E, Tobe H, Yamamoto K, Yamamoto S. Inhibition of angiogenesis by humulone, a bitter acid from beer hop. Biochem Biophys Res Commun, 2001, 289: 220–224

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education; School of Medicine, Xi’an Jiaotong University, Xi’an, 710061, China

    SiCen Wang, Meng Sun, YanMin Zhang, Jie Zhang & LangChong He

Authors
  1. SiCen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Meng Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. YanMin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jie Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. LangChong He
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to LangChong He.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, S., Sun, M., Zhang, Y. et al. EGFR/cell membrane chromatography-online-high performance liquid chromatography/mass spectrometry method for screening EGFR antagonists from Radix Angelicae Pubescentis . Sci. China Chem. 53, 2357–2362 (2010). https://doi.org/10.1007/s11426-010-4010-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11426-010-4010-3

Keywords

Search

Navigation