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Molecular recognition of small-cell lung cancer cells using aptamers.

ABSTRACT: Early diagnosis is the way to improve the rate of lung cancer survival, but is almost impossible today due to the lack of molecular probes that recognize lung cancer cells sensitively and selectively. We developed a new aptamer approach for the recognition of specific small-cell lung cancer (SCLC) cell-surface molecular markers. Our approach relies on cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX) to evolve aptamers for whole live cells that express a variety of surface markers representing molecular differences among cancer cells. When applied to different lung cancer cells including those from patient samples, these aptamers bind to SCLC cells with high affinity and specificity in various assay formats. When conjugated with magnetic and fluorescent nanoparticles, the aptamer nanoconjugates could effectively extract SCLC cells from mixed cell media for isolation, enrichment, and sensitive detection. These studies demonstrate the potential of the aptamer approach for early lung cancer detection.

SUBMITTER: Chen HW

PROVIDER: S-EPMC3544301 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature

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Molecular recognition of small-cell lung cancer cells using aptamers.

Chen Hui William HW Medley Colin D CD Sefah Kwame K Shangguan Dihua D Tang Zhiwen Z Meng Ling L Smith Josh E JE Tan Weihong W

ChemMedChem 20080601 6

Early diagnosis is the way to improve the rate of lung cancer survival, but is almost impossible today due to the lack of molecular probes that recognize lung cancer cells sensitively and selectively. We developed a new aptamer approach for the recognition of specific small-cell lung cancer (SCLC) cell-surface molecular markers. Our approach relies on cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX) to evolve aptamers for whole live cells that express a variety o ...[more]

PMID: 18338423

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Project description:AimTo generate DNA-aptamers binding to Methicillin-resistant Staphylococcus aureus (MRSA).MethodsThe Cell-Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology was used to run the selection against MRSA bacteria and develop target-specific aptamers. MRSA bacteria were targeted while Enterococcus faecalis bacteria were used for counter selection during that process. Binding assays to determine the right aptamer candidates as well as binding assays on clinical samples were performed through flow cytometry and analyzed using the FlowJo software. The characterization of the aptamers was done by determination of their Kd values and determined by analysis of flow data at different aptamer concentration using SigmaPlot. Finally, the recognition of the complex Gold-nanoparticle-aptamer to the bacteria cells was observed using transmission electron microscopy (TEM).ResultsDuring the cell-SELEX selection process, 17 rounds were necessary to generate enrichment of the pool. While the selection was run using fixed cells, it was shown that the binding of the pools with live cells was giving similar results. After sequencing and analysis of the two last pools, four sequences were identified to be aptamer candidates. The characterization of those aptamers showed that based on their Kd values, DTMRSA4 presented the best binding with a Kd value of 94.61 ± 18.82 nmol/L. A total of ten clinical samples of MRSA , S. aureus and Enterococcus faecalis were obtained to test those aptamers and determine their binding on a panel of samples. DTMRSA1 and DTMRSA3 showed the best results regarding their specificity to MRSA , DTMRSA1 being the most specific of all. Finally, those aptamers were coupled with gold-nanoparticle and their binding to MRSA cells was visualized through TEM showing that adduction of nanoparticles on the aptamers did not change their binding property.ConclusionA total of four aptamers that bind to MRSA were obtained with Kd values ranking from 94 to 200 nmol/L.

Dataset Information

Molecular recognition of small-cell lung cancer cells using aptamers.

Publications

Molecular recognition of small-cell lung cancer cells using aptamers.

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