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Updated: May 26, 2025
A Flow Cytometry-Based Cell Surface Protein Binding Assay for Assessing Selectivity and Specificity of an Anticancer Aptamer
Published on: September 13, 2022
Selection and identification of DNA aptamer binding VDAC1 for tumor tissue imaging and targeted drug delivery
Kai Zhang1, Baoyin Yuan1, Xiaoshuo Dai1
1Department of Pathophysiology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan Province 450001, PR China.
Abstract:
Hepatocellular carcinoma (HCC) represents a significant health concern. Identifying novel molecular targets is crucial for clinical diagnosis and targeted treatment of HCC. Aptamers are capable of binding specifically to cancer cells via target protein molecules. Consequently, aptamers are frequently employed to identify novel cancer biomarkers. The invasiveness of tumor cells is closely associated with the recurrence and metastasis of tumors. In this study, the highly invasive Huh7-P3 cells were initially constructed, and subsequently, several aptamers that could specifically recognize Huh7-P3 were developed using cell-based Systematic Evolution of Ligands by Exponential Enrichment (SELEX). The selected aptamer, designated S2-2, demonstrated the capacity to bind to multiple cancer cells. Furthermore, tissue imaging demonstrated that S2-2 exhibited a specific recognition of HCC tissue, while demonstrating no binding to normal tissue. Subsequently, voltage-dependent anion channel 1 (VDAC1) was identified as a potential target for S2-2. Furthermore, Doxorubicin (Dox)-loaded S2-2 was shown to specifically kill target Huh7-P3 cells. In vivo fluorescence imaging revealed that S2-2 was capable of specifically targeting tumors. Importantly, S2-2-Dox enhanced the anti-tumor efficacy of Dox in cell-line-derived xenograft (CDX) model. This study may provide a promising biomarker and molecular target for the clinical diagnosis and targeted therapy of cancers with high VDAC1 expression.
Insights
Researchers developed a novel aptamer, S2-2, that specifically targets hepatocellular carcinoma (HCC) cells and enhances chemotherapy. This aptamer shows promise as a biomarker and therapeutic agent for HCC and other cancers expressing VDAC1.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Hepatocellular carcinoma (HCC) poses a significant global health challenge.
- Novel molecular targets are essential for improving HCC diagnosis and treatment.
- Aptamers offer potential as specific cancer biomarkers and therapeutic delivery vehicles.
Purpose of the Study:
- To develop and characterize aptamers for specific recognition of highly invasive HCC cells.
- To identify the molecular target of the selected aptamer.
- To evaluate the therapeutic potential of aptamer-drug conjugates in preclinical models.
Main Methods:
- Cell-based Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was used to select aptamers against invasive HCC cells (Huh7-P3).
- Aptamer S2-2 was characterized for its binding specificity using tissue imaging and cell-based assays.
- Voltage-dependent anion channel 1 (VDAC1) was identified as the target of S2-2.
- Doxorubicin (Dox)-loaded S2-2 (S2-2-Dox) was tested for targeted cell killing and in vivo anti-tumor efficacy in a xenograft model.
Main Results:
- Aptamer S2-2 was successfully selected and demonstrated specific binding to HCC cells and tissues, with no significant binding to normal tissues.
- VDAC1 was confirmed as the molecular target of S2-2.
- S2-2-Dox exhibited specific killing of Huh7-P3 cells and enhanced anti-tumor efficacy in a preclinical HCC model.
- In vivo imaging confirmed S2-2's ability to specifically target tumors.
Conclusions:
- Aptamer S2-2 is a specific biomarker for HCC and targets VDAC1.
- S2-2-Dox demonstrates potent anti-tumor activity, suggesting its potential for targeted HCC therapy.
- This aptamer holds promise for clinical diagnosis and targeted treatment of cancers with high VDAC1 expression.
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