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DNA Aptamer Selected against Pancreatic Ductal Adenocarcinoma for in vivo Imaging and Clinical Tissue Recognition.

Wu X, Zhao Z, Bai H, Fu T, Yang C, Hu X, Liu Q, Champanhac C, Teng IT, Ye M, Tan W - Theranostics (2015)

Bottom Line: In this work, we have developed a truncated DNA aptamer, termed XQ-2d, with high affinity and specificity for pancreatic ductal adenocarcinoma (PDAC).Moreover, XQ-2d shows better recognition ratio for 40 tissue sections of clinical PDAC samples (82.5%) compared to the initial cell-SELEX selection library (5%).Therefore, XQ-2d can be considered a promising candidate as a tool for PDAC diagnosis and treatment.

View Article: PubMed Central - PubMed

Affiliation: 1. Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.

ABSTRACT
In this work, we have developed a truncated DNA aptamer, termed XQ-2d, with high affinity and specificity for pancreatic ductal adenocarcinoma (PDAC). Aptamer XQ-2d selectively binds to PL45 cells with a dissociation constant in the nanomolar range, as determined by its recognition of PL45 tumor cells in mice. Moreover, XQ-2d shows better recognition ratio for 40 tissue sections of clinical PDAC samples (82.5%) compared to the initial cell-SELEX selection library (5%). Therefore, XQ-2d can be considered a promising candidate as a tool for PDAC diagnosis and treatment.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric assay (a, b) and confocal microscopy assay (c) of the binding of aptamer XQ-2 (250 nM) to hTERT-HPNE cells and PL45 cells. The unselected initial library (250 nM) was used as control. Scale bar = 100 µm.
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Figure 3: Flow cytometric assay (a, b) and confocal microscopy assay (c) of the binding of aptamer XQ-2 (250 nM) to hTERT-HPNE cells and PL45 cells. The unselected initial library (250 nM) was used as control. Scale bar = 100 µm.

Mentions: After sequencing, the aptamer candidates were grouped based on homogeneity, and the first six groups were synthesized for further research. The results from flow cytometry and confocal microscopy imaging demonstrated that aptamer XQ-2 had good binding to the target PL45 cell line, but not to the negative cell line, hTERT-HPNE (Figure 3). These results suggested that aptamer XQ-2 might be capable of distinguishing PL45 cells from normal cells. Therefore, aptamer XQ-2 was further tested for its binding affinity to the PL45 cell line. To accomplish this, PL45 cells were incubated with different concentrations of FAM-labeled aptamers or initial library at 4 °C, followed by monitoring the fluorescence intensity of cell sample by flow cytometry. After the geometric mean fluorescence (GMF) intensity of cell samples treated with library was subtracted from that of cell samples treated with XQ-2, the equilibrium dissociation constants (Kd) of XQ-2 for PL45 cells were obtained by fitting the dependence of fluorescence intensity of samples on the concentration of the aptamers to the equation Y =B max X/( Kd +X). As shown in Supplementary Figure S1, the Kd of aptamer XQ-2 for PL45 cells was about 82.5 nM, demonstrating that the selected aptamer XQ-2 could bind with high affinity to target PL45 cells.


DNA Aptamer Selected against Pancreatic Ductal Adenocarcinoma for in vivo Imaging and Clinical Tissue Recognition.

Wu X, Zhao Z, Bai H, Fu T, Yang C, Hu X, Liu Q, Champanhac C, Teng IT, Ye M, Tan W - Theranostics (2015)

Flow cytometric assay (a, b) and confocal microscopy assay (c) of the binding of aptamer XQ-2 (250 nM) to hTERT-HPNE cells and PL45 cells. The unselected initial library (250 nM) was used as control. Scale bar = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4493536&req=5

Figure 3: Flow cytometric assay (a, b) and confocal microscopy assay (c) of the binding of aptamer XQ-2 (250 nM) to hTERT-HPNE cells and PL45 cells. The unselected initial library (250 nM) was used as control. Scale bar = 100 µm.
Mentions: After sequencing, the aptamer candidates were grouped based on homogeneity, and the first six groups were synthesized for further research. The results from flow cytometry and confocal microscopy imaging demonstrated that aptamer XQ-2 had good binding to the target PL45 cell line, but not to the negative cell line, hTERT-HPNE (Figure 3). These results suggested that aptamer XQ-2 might be capable of distinguishing PL45 cells from normal cells. Therefore, aptamer XQ-2 was further tested for its binding affinity to the PL45 cell line. To accomplish this, PL45 cells were incubated with different concentrations of FAM-labeled aptamers or initial library at 4 °C, followed by monitoring the fluorescence intensity of cell sample by flow cytometry. After the geometric mean fluorescence (GMF) intensity of cell samples treated with library was subtracted from that of cell samples treated with XQ-2, the equilibrium dissociation constants (Kd) of XQ-2 for PL45 cells were obtained by fitting the dependence of fluorescence intensity of samples on the concentration of the aptamers to the equation Y =B max X/( Kd +X). As shown in Supplementary Figure S1, the Kd of aptamer XQ-2 for PL45 cells was about 82.5 nM, demonstrating that the selected aptamer XQ-2 could bind with high affinity to target PL45 cells.

Bottom Line: In this work, we have developed a truncated DNA aptamer, termed XQ-2d, with high affinity and specificity for pancreatic ductal adenocarcinoma (PDAC).Moreover, XQ-2d shows better recognition ratio for 40 tissue sections of clinical PDAC samples (82.5%) compared to the initial cell-SELEX selection library (5%).Therefore, XQ-2d can be considered a promising candidate as a tool for PDAC diagnosis and treatment.

View Article: PubMed Central - PubMed

Affiliation: 1. Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.

ABSTRACT
In this work, we have developed a truncated DNA aptamer, termed XQ-2d, with high affinity and specificity for pancreatic ductal adenocarcinoma (PDAC). Aptamer XQ-2d selectively binds to PL45 cells with a dissociation constant in the nanomolar range, as determined by its recognition of PL45 tumor cells in mice. Moreover, XQ-2d shows better recognition ratio for 40 tissue sections of clinical PDAC samples (82.5%) compared to the initial cell-SELEX selection library (5%). Therefore, XQ-2d can be considered a promising candidate as a tool for PDAC diagnosis and treatment.

No MeSH data available.


Related in: MedlinePlus