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J. Wu



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    Poster Display Session (ID 63)

    • Event: ELCC 2017
    • Type: Poster Display Session
    • Track:
    • Presentations: 1
    • Moderators:
    • Coordinates: 5/07/2017, 12:30 - 13:00, Hall 1
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      33P - The screening and characterization of aptamer against gefitinib-resistant cells (ID 161)

      12:30 - 13:00  |  Author(s): J. Wu

      • Abstract

      Background:
      The EGFR tyrosine kinase inhibitor (TKI) gefitinib serves as first-line drug for patients with advanced non-small cell lung cancer (NSCLC) harboring EGFR mutation. The challenge of this target therapy is the acquired resistance due to T790M mutation after gefitinib treatment. Aptamers are single-strand DNA molecules that form 3D structures and specifically bind to target components. In this study, we wanted to isolate aptamers with recognition ability for gefitinib-resistant NSCLC cells and look for another mechanism that contributes to the acquired resistance that is not caused by T790M mutation.

      Methods:
      We used PC9 (gefitinib-sensitive cells) and PC9-IR (gefitinib-resistant cells with no T790M mutation) cells to select specific aptamers that bound to membrane proteins which were overexpressed in PC9-IR cells. We used suction-type microfluidic control module to perform cell-systematic evolution of ligands by exponential enrichment (Cell-SELEX) and obtained some aptamers that had more affinity to bind to PC9-IR. We examined the specificity of the aptamers and observed whether aptamers could bind to cell membranes of PC9-IR. In addition, we tested the cell cytotoxicity of the aptamers.

      Results:
      We obtained the aptamer, AP16-23F, which had greater affinity to bind to PC9-IR. In order to test whether AP16-23F could recognize the cells with gefitinib resistance, we used AP16-23F to isolate cells from PC9 by fluorescence-activated cell sorting (FACS). The results showed that the cells selected by AP16-23F were more resistant to gefitinib.

      Conclusions:
      We have isolated an aptamer with specificity for binding and capturing gefitinib-resistant NSCLC cells. This aptamer may be useful for drug resistance detection and may have the potential to deliver anti-cancer drug to gefitinib-resistant cells in the future.

      Clinical trial identification:


      Legal entity responsible for the study:
      National Cheng Kung University

      Funding:
      Ministry of Science and Technology, Taiwan

      Disclosure:
      All authors have declared no conflicts of interest.