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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

Scheme of systematic enrichment of aptamers against PDAC PL45 cell line. The DNA library was incubated with hTERT-HPNE cells (negative cells) to remove negative cell-binding sequences. The unbound DNAs were collected and then incubated with target PL45 cell line for positive selection. After washing, the bound DNAs were eluted and amplified by PCR for next-round selection. After enrichment, DNA sequencing was performed to identify individual aptamer sequences.
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Figure 1: Scheme of systematic enrichment of aptamers against PDAC PL45 cell line. The DNA library was incubated with hTERT-HPNE cells (negative cells) to remove negative cell-binding sequences. The unbound DNAs were collected and then incubated with target PL45 cell line for positive selection. After washing, the bound DNAs were eluted and amplified by PCR for next-round selection. After enrichment, DNA sequencing was performed to identify individual aptamer sequences.

Mentions: To generate aptamers against PDAC cancer cells, the human PDAC PL45 cell line was used as the target, and the normal hTERT-HPNE cell line was used for counter selection. The cell-SELEX process is schematically shown in Figure 1. For the first two rounds of selection, ssDNA library was only applied on PL45 cell monolayer for positive selection. From the third round of selection, the DNA library was first incubated with normal hTERT-HPNE cells to remove nonspecific sequences. The unbound DNA was collected and then incubated with target PL45 cells for positive selection. After washing, the bound DNA was eluted and amplified for next-round selection. During selection, the target cell-binding ability of evolved ssDNA library was monitored by flow cytometry. When PL45 cells were incubated with FAM-labeled ssDNA pools from an increasing number of selection rounds, steady increases in fluorescence intensity from the target cells were observed (Figure 2A). However, almost no increase in fluorescence signal was observed for hTERT-HPNE cells after incubation with FAM-labeled ssDNA pool from the 15th selection round (Figure 2B). Therefore, the ssDNA pool evolved from the 15th round of selection was high-throughput sequenced with Illumina MiSeq.


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)

Scheme of systematic enrichment of aptamers against PDAC PL45 cell line. The DNA library was incubated with hTERT-HPNE cells (negative cells) to remove negative cell-binding sequences. The unbound DNAs were collected and then incubated with target PL45 cell line for positive selection. After washing, the bound DNAs were eluted and amplified by PCR for next-round selection. After enrichment, DNA sequencing was performed to identify individual aptamer sequences.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Scheme of systematic enrichment of aptamers against PDAC PL45 cell line. The DNA library was incubated with hTERT-HPNE cells (negative cells) to remove negative cell-binding sequences. The unbound DNAs were collected and then incubated with target PL45 cell line for positive selection. After washing, the bound DNAs were eluted and amplified by PCR for next-round selection. After enrichment, DNA sequencing was performed to identify individual aptamer sequences.
Mentions: To generate aptamers against PDAC cancer cells, the human PDAC PL45 cell line was used as the target, and the normal hTERT-HPNE cell line was used for counter selection. The cell-SELEX process is schematically shown in Figure 1. For the first two rounds of selection, ssDNA library was only applied on PL45 cell monolayer for positive selection. From the third round of selection, the DNA library was first incubated with normal hTERT-HPNE cells to remove nonspecific sequences. The unbound DNA was collected and then incubated with target PL45 cells for positive selection. After washing, the bound DNA was eluted and amplified for next-round selection. During selection, the target cell-binding ability of evolved ssDNA library was monitored by flow cytometry. When PL45 cells were incubated with FAM-labeled ssDNA pools from an increasing number of selection rounds, steady increases in fluorescence intensity from the target cells were observed (Figure 2A). However, almost no increase in fluorescence signal was observed for hTERT-HPNE cells after incubation with FAM-labeled ssDNA pool from the 15th selection round (Figure 2B). Therefore, the ssDNA pool evolved from the 15th round of selection was high-throughput sequenced with Illumina MiSeq.

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