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

November-2014 Volume 32 Issue 5

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

November-2014 Volume 32 Issue 5

Full Size Image

Article

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
View Affiliations / Copyright

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
Pages: 2054-2060
|
Published online on: August 18, 2014

https://doi.org/10.3892/or.2014.3411
Expand metrics +

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.

View Figures

View References

1	Pietrantonio F, De Braud F, Da Prat V, et al: A review on biomarkers for prediction of treatment outcome in gastric cancer. Anticancer Res. 33:1257–1266. 2013.PubMed/NCBI
2	Takahashi T, Saikawa Y and Kitagawa Y: Gastric cancer: current status of diagnosis and treatment. Cancers. 5:48–63. 2013. View Article : Google Scholar : PubMed/NCBI
3	Jemal A, Bray F, Center MM, et al: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
4	Yang S and Chung HC: Novel biomarker candidates for gastric cancer. Oncol Rep. 19:675–680. 2008.PubMed/NCBI
5	Lassalle HP, Marchal S, Guillemin F, et al: Aptamers as remarkable diagnostic and therapeutic agents in cancer treatment. Curr Drug Metab. 13:1130–1144. 2012. View Article : Google Scholar : PubMed/NCBI
6	Radom F, Jurek PM, Mazurek MP, et al: Aptamers: molecules of great potential. Biotechnol Adv. 31:1260–1274. 2013. View Article : Google Scholar : PubMed/NCBI
7	Hermann T and Patel DJ: Adaptive recognition by nucleic acid aptamers. Science. 287:820–825. 2000. View Article : Google Scholar : PubMed/NCBI
8	Tuerk C and Gold L: Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science. 249:505–510. 1990. View Article : Google Scholar
9	Stoltenburg R, Reinemann C and Strehlitz B: SELEX - a (r)evolutionary method to generate high-affinity nucleic acid ligands. Biomol Eng. 24:381–403. 2007. View Article : Google Scholar : PubMed/NCBI
10	Sefah K, Shangguan D, Xiong X, et al: Development of DNA aptamers using Cell-SELEX. Nat Protoc. 5:1169–1185. 2010. View Article : Google Scholar : PubMed/NCBI
11	Cerchia L and de Franciscis V: Targeting cancer cells with nucleic acid aptamers. Trends Biotechnol. 28:517–525. 2010. View Article : Google Scholar : PubMed/NCBI
12	Shangguan D, Li Y, Tang Z, et al: Aptamers evolved from live cells as effective molecular probes for cancer study. Proc Natl Acad Sci USA. 103:11838–11843. 2006. View Article : Google Scholar : PubMed/NCBI
13	Sefah K, Tang ZW, Shangguan DH, et al: Molecular recognition of acute myeloid leukemia using aptamers. Leukemia. 23:235–244. 2009. View Article : Google Scholar : PubMed/NCBI
14	Zhang K, Sefah K, Tang L, et al: A novel aptamer developed for breast cancer cell internalization. Chem Med Chem. 7:79–84. 2012. View Article : Google Scholar : PubMed/NCBI
15	Kunii T, Ogura S, Mie M and Kobatake E: Selection of DNA aptamers recognizing small cell lung cancer using living cell-SELEX. Analyst. 136:1310–1312. 2011. View Article : Google Scholar : PubMed/NCBI
16	Sefah K, Meng L, Lopez-Colon D, et al: DNA aptamers as molecular probes for colorectal cancer study. PLoS One. 5:e142692010. View Article : Google Scholar : PubMed/NCBI
17	Zhu G, Ye M, Donovan MJ, et al: Nucleic acid aptamers: an emerging frontier in cancer therapy. Chem Commun. 48:10472–10480. 2012. View Article : Google Scholar : PubMed/NCBI
18	Zhang Y, Chen Y, Han D, et al: Aptamers selected by cell-SELEX for application in cancer studies. Bioanalysis. 2:907–918. 2010. View Article : Google Scholar : PubMed/NCBI
19	Sefah K, Bae KM, Phillips JA, et al: Cell-based selection provides novel molecular probes for cancer stem cells. Int J Cancer. 132:2578–2588. 2013. View Article : Google Scholar : PubMed/NCBI
20	Zueva E, Rubio LI, Ducongé F and Tavitian B: Metastasis-focused cell-based SELEX generates aptamers inhibiting cell migration and invasion. Int J Cancer. 128:797–804. 2011. View Article : Google Scholar : PubMed/NCBI
21	Berezovski MV, Lechmann M, Musheev MU, et al: Aptamer-facilitated biomarker discovery (AptaBiD). J Am Chem Soc. 130:9137–9143. 2008. View Article : Google Scholar : PubMed/NCBI
22	Fang X and Tan W: Aptamers generated from cell-SELEX for molecular medicine: a chemical biology approach. Acc Chem Res. 43:48–57. 2009. View Article : Google Scholar : PubMed/NCBI
23	Pavlou MP, Diamandis EP and Blasutig IM: The long journey of cancer biomarkers from the bench to the clinic. Clin Chem. 59:147–157. 2013. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

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

This article is mentioned in:

Abstract

Figure 1

Figure 2

Figure 3

Figure 4

Related Articles