Selection of DNA aptamers against glioblastoma cells with high affinity and specificity

PLoS One. 2012;7(10):e42731. doi: 10.1371/journal.pone.0042731. Epub 2012 Oct 2.

Abstract

Background: Glioblastoma is the most common and most lethal form of brain tumor in human. Unfortunately, there is still no effective therapy to this fatal disease and the median survival is generally less than one year from the time of diagnosis. Discovery of ligands that can bind specifically to this type of tumor cells will be of great significance to develop early molecular imaging, targeted delivery and guided surgery methods to battle this type of brain tumor.

Methodology/principal findings: We discovered two target-specific aptamers named GBM128 and GBM131 against cultured human glioblastoma cell line U118-MG after 30 rounds selection by a method called cell-based Systematic Evolution of Ligands by EXponential enrichment (cell-SELEX). These two aptamers have high affinity and specificity against target glioblastoma cells. They neither recognize normal astraglial cells, nor do they recognize other normal and cancer cell lines tested. Clinical tissues were also tested and the results showed that these two aptamers can bind to different clinical glioma tissues but not normal brain tissues. More importantly, binding affinity and selectivity of these two aptamers were retained in complicated biological environment.

Conclusion/significance: The selected aptamers could be used to identify specific glioblastoma biomarkers. Methods of molecular imaging, targeted drug delivery, ligand guided surgery can be further developed based on these ligands for early detection, targeted therapy, and guided surgery of glioblastoma leading to effective treatment of glioblastoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism*
  • Astrocytoma / chemistry
  • Astrocytoma / genetics
  • Astrocytoma / metabolism
  • Base Sequence
  • Binding, Competitive
  • Brain Neoplasms / metabolism*
  • Brain Neoplasms / pathology
  • Carbocyanines / chemistry
  • Carbocyanines / metabolism
  • Cell Line
  • Cell Line, Tumor
  • Flow Cytometry
  • Glioblastoma / metabolism
  • Glioblastoma / pathology
  • HEK293 Cells
  • HT29 Cells
  • HeLa Cells
  • Humans
  • MCF-7 Cells
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Oligodendroglioma / chemistry
  • Oligodendroglioma / genetics
  • Oligodendroglioma / metabolism
  • SELEX Aptamer Technique / methods*

Substances

  • Aptamers, Nucleotide
  • Carbocyanines
  • cyanine dye 5

Grants and funding

This work was supported by the National Basic Research Program of China (2010CB732402), National Instrumentation Program (2011YQ03012412), the Natural Science Foundation of Fujian Province for Distinguished Young Scholars (2010 J06004), and the Key Laboratory for Chemical Biology of Fujian Province. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.