Quantitative Multi-targeted Imaging of Metastatic Colorectal Cancer Cells Using Aptamer Probes in Combination†

doi:10.7503/cjcu20160254

Abstract

Abstract:

We developed seven aptamer probes towards metastatic colorectal cancer cells by conjugating the aptamers to quantum dots(QD605) for multi-targeted cancer cell imaging. The results of competitive assay showed that the seven aptamers could recognize their individual targets on the same cell without any mutual interference, revealing that they may be applied in combination for multi-target cancer cell imaging. The results of quantitative assay of the fluorescence images showed that compared with a single probe, the seven probes in combination can significantly increase the fluorescent intensity on the cell surface, and the detection rate of positive cells increased significantly, resulting in a higher detection sensitivity. Further, the probes were used to detect several colorectal cancer cell lines using flow cytometry. The results showed that seven probes used in combination could effectively evaluate colorectal cancer metastatic potential. This study demonstrated that usage of multiple aptamer probes in combination could effectively improve the sensitivity and accuracy of target imaging which provides a novel approach for targeted diagnosis of colorectal cancer.

Key words: Aptamer, Metastatic colorectal cancer, Multi-target, Quantum dot, Molecular imaging

CLC Number:

O657.3

TrendMD:

LI Wanming, FANG Jin. Quantitative Multi-targeted Imaging of Metastatic Colorectal Cancer Cells Using Aptamer Probes in Combination^†[J]. Chem. J. Chinese Universities, 2016, 37(7): 1262.

Figures/Tables 8

Table 1 DNA sequences used in experiments

DNA probe	Sequence(5-3)
Library	AGGAGCAGCGTGGAGGATA(45N)TTAGGGTGTGTCGTCGTGGT
W1	ATTGATGACACTGAATTGATTTCAGGCATTGTTTGCTGCCACTTAGGGTGTGTCGTCG
W3	AGCAGCGTGGAGGATAGGGGTCGGAGTGGGTGGTTATGATTGGCTCTTCTGCGCTGC
W6	ACCCTATCGGTTTCTTGTCTTGCAAGCCTTCAGCTTCAGCAGCCTTAGGG
W9	ACGCAACCTCTTGGCTCATTGACTGAGGCCATTGTTTGTTGGCTTAGGGTGTG
W14	AGGATACAGGTTGGCTTTGGTGGCTATGGGTTGGGTGTGGTGCGTATCCT
W17	AGGAGCAGCGTGGAGGATATGATCCGACCGTTTTCGTTTGTTATGAACGGTGCTCC
W22	ACAGCGTGGAGGATATTGGTGTTGGTGGGCGTTTACGCTCCTTCGTCGCTG

Fig.1 Flow cytometry for the binding of FITC-labeled aptamers to LoVo(A1—A7) and HCT-8(B1—B7) cellsa. Library; b. aptamer. (A1, B1) W1; (A2, B2) W3; (A3, B3) W6; (A4, B4) W9; (A5, B5) W14; (A6, B6) W17; (A7, B7) W22.

Fig.2 Quantitative results based on flow cytometry Data were presented as the mean±standard deviation.

Fig.3 Mutual effects of aptamers on the recognition of targets by competitive experiment(A) FITC-W1; (B) FITC-W3; (C) FITC-W6; (D) FITC-W9; (E) FITC-W14; (F) FITC-W17; (G) FITC-W22.

Fig.4 Fluorescence(A1—A9) and optical(B1—B9) images of LoVo cells stained by aptamers-QDAll pictures were taken under a fluorescence microscope with × 200 magnification.(A1, B1) Library; (A2, B2) W1; (A3, B3) W3; (A4, B4) W6; (A5, B5) W9; (A6, B6) W14; (A7, B7) W17; (A8, B8) W22; (A9, B9) multiple aptamers.

Fig.5 Quantitative results based on QD-cellsa. Library; b. W1; c. W3; d. W6; e. W9; f. W14; g. W17; h. W22; i. multiple aptamers.

Fig.6 Flow cytometry assays of the binding of aptamers with cells (A) LoVo; (B) HCT116; (C) SW620; (D) CL187; (E) HEK293.

Fig.7 Relative fluorescence intensities of aptamers-cells

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