Evolution of a gastric carcinoma cell-specific DNA aptamer by live cell-SELEX

Cao,Hong-Yong; Yuan,Ai-Hua; Shi,Xue-Song; Chen,Wei; Miao,Yi

doi:10.3892/or.2014.3411

Evolution of a gastric carcinoma cell-specific DNA aptamer by live cell-SELEX

Authors:
- Hong-Yong Cao
- Ai-Hua Yuan
- Xue-Song Shi
- Wei Chen
- Yi Miao
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Affiliations: Department of General Surgery, Nanjing Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu, P.R. China, Department of General Surgery, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, P.R. China
Published online on: August 18, 2014 https://doi.org/10.3892/or.2014.3411
Pages: 2054-2060

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Abstract

Aptamers have emerged as promising molecular probes for disease diagnosis and therapy. In the present study, the entire live cell-SELEX method was used to generate gastric cancer cell‑specific aptamers. Human gastric carcinoma AGS cells were used as target cells for positive selections and human normal gastric epithelial GES-1 cells as the negative cells for counter selections. The selection procedure was monitored by gel electrophoresis and flow cytometric analysis. By successive in vitro evolutions and subsequent cloning and sequencing, a gastric carcinoma cell‑specific DNA aptamer termed cy-apt 20 with minimal recognition to the controls was identified from the final enriched ssDNA pool. Flow cytometry binding assays revealed that cy-apt 20 had a >70% binding rate to AGS cells and <30% binding affinity to non-gastric cancer cells. Furthermore, the targeting recognition of AGS cells was established by using minimal doses of FITC-cy-apt 20 that continued for a long period of time. As visualized by fluorescence imaging, the majority of AGS cells were stained by FITC-cy-apt 20. The fluorescence intensity of AGS cells was ~6-fold over that of non-AGS cells. The present study demonstrated that the entire live cell-SELEX was simple, but effective in generating gastric cancer cell‑specific aptamers, and that the aptamer cy-apt 20 has great potential to be used for the study and diagnosis of gastric cancer.

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