In vitro selection of aptamer S1 against MCF-7 human breast cancer cells |
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| Institution: | 1. Department of Pharmacy, Xiamen Medical College, Xiamen 361023, China;2. School of Pharmaceutical Sciences, Xiamen University, Xiamen, Fujian 361102, China;3. Fujian Provincial Key Laboratory of Innovative Drug Target Research, Xiamen, Fujian 361102, China;1. Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou, 510006, China;2. Guangzhou Key Laboratory of Construction and Application of New Drug Screening Model Systems, Guangdong Pharmaceutical University, Guangzhou, 510006, China;3. Key Laboratory of New Drug Discovery and Evaluation of Ordinary Universities of Guangdong Province, Guangdong Pharmaceutical University, Guangzhou, 510006, China;4. School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, 510006, China;1. Division of Cancer Biology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan;2. Laboratory of Histology and Cytology, Hokkaido University Graduate School of Medicine, Sapporo, Japan;3. Chemistry Department, Faculty of Science, Tanta University, Egypt;4. National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA;5. Molsys Scientific, 01 Kogilu, Mittiganahalli cross Yelahanka, Bangalore, 560064, India;1. Department of Gastrointestinal Surgery, Tongji Hospital, Huazhong University of Science and Technology, No. 1095 Jiefang Avenue, Wuhan, Hubei Province, China;2. Department of Obstetrics and Gynecology, Tongji Hospital, Huazhong University of Science and Technology, No. 1095 Jiefang Avenue, Wuhan, Hubei Province, China;3. Cancer Research Institute, Tongji Hospital, Huazhong University of Science and Technology, No. 1095 Jiefang Avenue, Wuhan, Hubei Province, China;1. CNRS, UMR 7242, Biotechnologie et Signalisation Cellulaire, Institut de Recherche de l’Ecole de Biotechnologie de Strasbourg, Université de Strasbourg, 67400 Illkirch-Graffenstaden, France;2. CNRS, UMR 7021, Laboratoire de Bioimagerie et Pathologies, Tumoral Signaling and Therapeutic Targets, Faculté de Pharmacie, Université de Strasbourg, 67401 Illkirch, France;3. Département de Pharmacie, Centre de Lutte Contre le Cancer Paul Strauss, 67000 Strasbourg, France;4. EA 3430, Progression Tumorale et Micro-environnement, Approches Translationnelles et Épidémiologie, Université de Strasbourg, 67000 Strasbourg, France;5. CNRS, UMR 7199, Laboratoire de Conception et Application de Molécules Bioactives, Faculté de Pharmacie, Université de Strasbourg, 67401 Illkirch, France |
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| Abstract: | Breast cancer is the most common female cancer. However, the known effective specific biomarkers for breast cancer are still scarce. Abnormal membrane proteins serve as ideal biomarkers for disease diagnoses, therapeutics and prognosis. Thus aptamers (single-stranded oligonucleotide molecules) with molecular recognition properties can be used as efficient tools to sort cells based on differences in cell surface architecture between normal and tumor cells. In this study, we aimed to screen specific aptamer against MCF-7 human breast cancer cells. Cell-SELEX process was performed to isolate aptamers from a combinatorial single-stranded nucleic acid library that selectively targeting surface proteins of MCF-7 cells in contrast with MCF-10A human mammary epithelial cells. The process was repeated until the pool was enriched for sequences that specifically recognizing MCF-7 cells in monitoring by flow cytometry. Subsequently, the enriched pool was cloned into bacteria, and positive clones were sequenced to obtain individual sequences. Representative sequences were chemically synthesized and evaluated their binding affinities to MCF-7 cells. As a result, an aptamer S1 was finally identified to have high binding affinity with equilibrium dissociation constant (Kd) value of 29.9 ± 6.0 nM. FAM-labeled aptamer S1 induced fluorescence shift in MCF-7 cells but not in MCF-10A human mammary epithelial cells, or MDA-MB-453 and MDA-MB-231 human breast cancer cells. Furthermore, result of cell imaging observed from laser confocal fluorescence microscope showed that MCF-7 cells exhibited stronger fluorescence signal resulted from Cy5-labeled aptamer S1 than MCF-10A cells. The above findings suggested that S1 may be a specificity and selectivity aptamer for MCF-7 cells and useful for the breast cancer detection and diagnosis. |
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| Keywords: | Breast cancer Selection Aptamer Disease diagnoses MCF-7 cell |
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