Open Access

Application of a small molecule inhibitor screen approach to identify CXCR4 downstream signaling pathways that promote a mesenchymal and fulvestrant‑resistant phenotype in breast cancer cells

  • Authors:
    • Margarite D. Matossian
    • Steven Elliott
    • Lyndsay V. Rhodes
    • Elizabeth  C. Martin
    • Van T. Hoang
    • Hope E. Burks
    • William J. Zuercher
    • David  H. Drewry
    • Bridgette M. Collins‑Burow
    • Matthew E. Burow
  • View Affiliations

  • Published online on: March 16, 2021     https://doi.org/10.3892/ol.2021.12641
  • Article Number: 380
  • Copyright: © Matossian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Chemokine receptor 4 (CXCR4) and its ligand stromal‑derived factor 1 (SDF‑1) have well‑characterized functions in cancer metastasis; however, the specific mechanisms through which CXCR4 promotes a metastatic and drug‑resistant phenotype remain widely unknown. The aim of the present study was to demonstrate the application of a phenotypic screening approach using a small molecule inhibitor library to identify potential CXCR4‑mediated signaling pathways. The present study demonstrated a new application of the Published Kinase Inhibitor Set (PKIS), a library of small molecule inhibitors from diverse chemotype series with varying levels of selectivity, in a phenotypic medium‑throughput screen to identify potential mechanisms to pursue. Crystal violet staining and brightfield microscopy were employed to evaluate relative cell survival and changes to cell morphology in the screens. ‘Hits’ or lead active compounds in the first screen were PKIS inhibitors that reversed mesenchymal morphologies in CXCR4‑activated breast cancer cells without the COOH‑terminal domain (MCF‑7‑CXCR4‑ΔCTD) and in the phenotypically mesenchymal triple‑negative breast cancer cells (MDA‑MB‑231, BT‑549 and MDA‑MB‑157), used as positive controls. In a following screen, the phenotypic and cell viability screen was used with a positive control that was both morphologically mesenchymal and had acquired fulvestrant resistance. Compounds within the same chemotype series were identified that exhibited biological activity in the screens, the ‘active’ inhibitors, were compared with inactive compounds. Relative kinase activity was obtained using published datasets to discover candidate kinase targets responsible for CXCR4 activity. MAP4K4 and MINK reversed both the mesenchymal and drug‑resistant phenotypes, NEK9 and DYRK2 only reversed the mesenchymal morphology, and kinases, including ROS, LCK, HCK and LTK, altered the fulvestrant‑resistant phenotype. Oligoarray experiments revealed pathways affected in CXCR4‑activated cells, and these pathways were compared with the present screening approach to validate our screening tool. The oligoarray approach identified the integrin‑mediated, ephrin B‑related, RhoA, RAC1 and ErbB signaling pathways to be upregulated in MCF‑7‑CXCR4‑ΔCTD cells, with ephrin B signaling also identified in the PKIS phenotypic screen. The present screening tool may be used to discover potential mechanisms of targeted signaling pathways in solid cancers.
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May-2021
Volume 21 Issue 5

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Spandidos Publications style
Matossian MD, Elliott S, Rhodes LV, Martin EC, Hoang VT, Burks HE, Zuercher WJ, Drewry DH, Collins‑Burow BM, Burow ME, Burow ME, et al: Application of a small molecule inhibitor screen approach to identify CXCR4 downstream signaling pathways that promote a mesenchymal and fulvestrant‑resistant phenotype in breast cancer cells. Oncol Lett 21: 380, 2021.
APA
Matossian, M.D., Elliott, S., Rhodes, L.V., Martin, E.C., Hoang, V.T., Burks, H.E. ... Burow, M.E. (2021). Application of a small molecule inhibitor screen approach to identify CXCR4 downstream signaling pathways that promote a mesenchymal and fulvestrant‑resistant phenotype in breast cancer cells. Oncology Letters, 21, 380. https://doi.org/10.3892/ol.2021.12641
MLA
Matossian, M. D., Elliott, S., Rhodes, L. V., Martin, E. C., Hoang, V. T., Burks, H. E., Zuercher, W. J., Drewry, D. H., Collins‑Burow, B. M., Burow, M. E."Application of a small molecule inhibitor screen approach to identify CXCR4 downstream signaling pathways that promote a mesenchymal and fulvestrant‑resistant phenotype in breast cancer cells". Oncology Letters 21.5 (2021): 380.
Chicago
Matossian, M. D., Elliott, S., Rhodes, L. V., Martin, E. C., Hoang, V. T., Burks, H. E., Zuercher, W. J., Drewry, D. H., Collins‑Burow, B. M., Burow, M. E."Application of a small molecule inhibitor screen approach to identify CXCR4 downstream signaling pathways that promote a mesenchymal and fulvestrant‑resistant phenotype in breast cancer cells". Oncology Letters 21, no. 5 (2021): 380. https://doi.org/10.3892/ol.2021.12641