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In vitro selection of DNA aptamers against human osteosarcoma
Khaliunsarnai Tsogtbaatar1,2, Diana A Sousa2, Debora Ferreira2
1Institute of Science, Hacettepe University, Bioengineering Division, 06800, Ankara, Turkey.
Background:
Osteosarcoma is a highly malignant bone tumor, most frequently occurring in the rapid bone growth phase. Effective treatment of this disease is hindered by the lack of specific probes for early diagnosis and the fast cancer widespread.
Methods:
To find such probes, the cell-Systematic Evolution of Ligands by EXponential enrichment (cell-SELEX) methodology was implemented against the human osteosarcoma MG-63 cell line towards the selection of new specific aptamers. After 10 rounds of selection, the aptamer DNA pool was Sanger sequenced and the sequences were subjected to a bioinformatic analysis that included sequence alignment, phylogenetic relationship, and secondary structure prediction.
Results:
A DNA aptamer (OS-7.9), with a dissociation constant (Kd) value in the nanomolar range (12.8 ± 0.9 nM), revealed high affinity against the target cells at the physiological temperature. Furthermore, the selected aptamer also recognized lung carcinoma and colon colorectal adenocarcinoma cell lines, which are reported as common metastasis sites of osteosarcoma.
Conclusions:
These results suggest that OS-7.9 could recognize a common protein expressed in these cancer cells, possibly becoming a potential molecular probe for early diagnosis and targeted therapies for metastatic disease. Moreover, to the best of our knowledge, this was the first attempt to generate a DNA aptamer (OS-7.9 aptamer) against the MG-63-cell line by cell-SELEX.
Insights
Researchers developed a novel DNA aptamer (OS-7.9) using cell-SELEX to detect osteosarcoma. This aptamer shows high affinity for cancer cells, offering potential for early diagnosis and targeted therapy of metastatic disease.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Osteosarcoma is a highly malignant bone tumor.
- Early diagnosis and treatment are challenged by rapid cancer spread.
- Lack of specific probes hinders effective management.
Purpose of the Study:
- To develop specific DNA aptamers for osteosarcoma detection.
- To identify molecular probes for early diagnosis and targeted therapy.
- To address the need for improved diagnostic tools in osteosarcoma.
Main Methods:
- Implemented cell-Systematic Evolution of Ligands by EXponential enrichment (cell-SELEX) methodology.
- Utilized human osteosarcoma MG-63 cell line for aptamer selection.
- Performed Sanger sequencing and bioinformatic analysis of selected aptamer sequences.
Main Results:
- Identified a DNA aptamer (OS-7.9) with high affinity (Kd = 12.8 ± 0.9 nM) for target cells at physiological temperature.
- OS-7.9 aptamer also recognized lung carcinoma and colorectal adenocarcinoma cell lines.
- Demonstrated the aptamer's potential to bind common proteins on metastatic cancer cells.
Conclusions:
- OS-7.9 aptamer shows promise as a molecular probe for early osteosarcoma diagnosis.
- The aptamer could be valuable for targeted therapies in metastatic osteosarcoma.
- This represents the first DNA aptamer generated against MG-63 cells using cell-SELEX.
