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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

(a) Binding ability of aptamer XQ-2 (250 nM) to PL45 cells with no pretreatment and pretreatment with proteinase K or trypsin for 10 min. (b) Internalized assay of aptamer XQ-2 (250 nM) in PL45 cells after incubation at 37 °C for 2 h.
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Figure 5: (a) Binding ability of aptamer XQ-2 (250 nM) to PL45 cells with no pretreatment and pretreatment with proteinase K or trypsin for 10 min. (b) Internalized assay of aptamer XQ-2 (250 nM) in PL45 cells after incubation at 37 °C for 2 h.

Mentions: Because of its overexpression in certain kinds of cancer cell lines, the target of aptamer XQ-2 was further characterized. To determine if the binding target of XQ-2 is an extracellular membrane protein, the binding affinity of XQ-2 against PL45 cells treated with trypsin or proteinase K was investigated. For PL45 cells treated with trypsin or proteinase K for 10 min, as shown in Figure 5A, XQ-2 lost its binding ability. The significant loss of binding ability of XQ-2 to cells treated with proteinase suggests that the binding targets of XQ-2 are most likely extracellular proteins. To investigate whether XQ-2 could be internalized into PL45 cells, the intracellular fluorescence signals of the targeted cells were analyzed by flow cytometry after removing the fluorescence signals on the cell surface by trypsin or proteinase K. As shown in Figure 5B, the removal of the fluorescence signals on the cell surface caused the disappearance of fluorescence signals of target cells, indicating there were little intracellular fluorescence signals. Thus, it might be deduced from these data that aptamer XQ-2 mainly localized on the target PL45 cell surface after incubation at 37 °C for 2 h.


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)

(a) Binding ability of aptamer XQ-2 (250 nM) to PL45 cells with no pretreatment and pretreatment with proteinase K or trypsin for 10 min. (b) Internalized assay of aptamer XQ-2 (250 nM) in PL45 cells after incubation at 37 °C for 2 h.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493536&req=5

Figure 5: (a) Binding ability of aptamer XQ-2 (250 nM) to PL45 cells with no pretreatment and pretreatment with proteinase K or trypsin for 10 min. (b) Internalized assay of aptamer XQ-2 (250 nM) in PL45 cells after incubation at 37 °C for 2 h.
Mentions: Because of its overexpression in certain kinds of cancer cell lines, the target of aptamer XQ-2 was further characterized. To determine if the binding target of XQ-2 is an extracellular membrane protein, the binding affinity of XQ-2 against PL45 cells treated with trypsin or proteinase K was investigated. For PL45 cells treated with trypsin or proteinase K for 10 min, as shown in Figure 5A, XQ-2 lost its binding ability. The significant loss of binding ability of XQ-2 to cells treated with proteinase suggests that the binding targets of XQ-2 are most likely extracellular proteins. To investigate whether XQ-2 could be internalized into PL45 cells, the intracellular fluorescence signals of the targeted cells were analyzed by flow cytometry after removing the fluorescence signals on the cell surface by trypsin or proteinase K. As shown in Figure 5B, the removal of the fluorescence signals on the cell surface caused the disappearance of fluorescence signals of target cells, indicating there were little intracellular fluorescence signals. Thus, it might be deduced from these data that aptamer XQ-2 mainly localized on the target PL45 cell surface after incubation at 37 °C for 2 h.

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