Cancer diagnosis and treatment integrated nano reagent as well as preparation method and application thereof

A reagent and cancer technology, applied in the field of cancer diagnosis and treatment integrated nano reagents and its preparation, to achieve the effect of SERS signal enhancement

Active Publication Date: 2022-06-03
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, methods to achieve intercellular miRNA-triggered DNAzyme-based dual gene silencing, especially an all-in-one reagent for synergistic enhancement of target-triggered nanoparticle cascade assembly and DNAzyme amplification for cancer diagnosis and therapy have not yet been reported.

Method used

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  • Cancer diagnosis and treatment integrated nano reagent as well as preparation method and application thereof
  • Cancer diagnosis and treatment integrated nano reagent as well as preparation method and application thereof
  • Cancer diagnosis and treatment integrated nano reagent as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation of miRNA-responsive cancer diagnosis and treatment integrated reagents

1. Preparation of AuNP-Y

1) Mix equal amounts (50 μM, 10 μL) of Y-motif ssDNA-A, Y-motifssDNA-B and Y-motif ssDNA-C (i.e. Ya, Yb and Yc, respectively) in 50 μL PBS buffer, Then annealing is performed to assemble the Y-shaped DNA structure, that is, the Y-motif;

2) After incubating 50 μL solution containing the above 10 μM Y-motif with 500 μL gold nanoparticles (0.3 nM), that is, AuNPs at room temperature for 12 h, the aging treatment was performed, that is, sodium chloride solution was added four times, each time At 30 min intervals, the final sodium chloride concentration was 0.2 M, and the mixture was gently shaken at room temperature for 12 h;

3) Finally, wash with PBS three times by centrifugation at a rotational speed of 7000 rpm, and the time of each centrifugation is 20 min; then resuspend the precipitate in 50 μL of PBS to obtain AuNP-Y, and store at 4°C for fu...

Embodiment 2

[0024] Example 2: Reaction mechanism characterization of miRNA-responsive cancer diagnosis and treatment integrated reagents

[0025] figure 2The formation of Y-motif and its specific response to miRNA-106a and ATP were characterized by 10% PAGE in . The DNA samples correspond specifically to DNA samples in different lanes, namely: M: DNA marker; Lane 1: Ya; Lane 2: Yb; Lane 3: Yc; Lane 4: Ya, Yb and Yc in equal amounts Hybridization (Y); Lane 5: miRNA-106a; Lane 6: Equivalent hybridization of Y with miRNA-106a; Lane 7: Equivalent hybridization of Yb with Yc; Lane 8: L; Lane 9: c-Jun; Lane 10: L Equal amounts of hybridization with c-Jun; Lane 11: Yb and Yc hybridization products react with equal amounts of L and c-Jun hybridization products; Lane 12: Equal amounts of Yc and L hybridization; Lane 13: Y, miRNA-106a, ATP and L Equivalent hybridization with the c-Jun hybridization product. The main band of the hybridization product in lane 4 electrophoresed slower than the t...

Embodiment 3

[0027] Example 3: Feasibility verification of miRNA-triggered AuNP networked nanostructure assembly and SERS sensing

An equal amount (10 μL) of AuNP-Y and AuNP-D in Example 1 was mixed with 80 μL PBS containing 0.5 nM miRNA-106a, and incubated at 37°C with gentle shaking for 2 h to form target-induced AuNP networked nanoparticles. structure. The AuNP network nanostructures were characterized by absorption, dynamic light scattering (DLS) and scanning electron microscopy (SEM).

[0028] like Figure 4A As shown, the surface plasmon resonance (SPR) peak of AuNPs was red-shifted from 524 nm to 526.4 nm by the modification of Y-motifs. Similarly, through the modification of dsDNA linker and DTNB, the SPR of AuNPs was red-shifted to 526 nm, which indicated that AuNP-Ds were successfully prepared.

[0029] like Figure 4B The DLS characterization shown, the hydrodynamic diameters of AuNP-Y and AuNP-D are about 33 nm and 31.6 nm, respectively, and the SEM images show that AuNP-...

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Abstract

The invention discloses a cancer diagnosis and treatment integrated nano reagent which comprises a miRNA recognition probe prepared by modifying the surface of AuNP with a Y-type DNA structure and an SERS probe prepared by modifying AuNP with a double-stranded DNA connector and Raman molecules, and the invention further discloses a preparation method and application of the cancer diagnosis and treatment integrated reagent. According to the cancer diagnosis and treatment integrated nano reagent disclosed by the invention, under the driving of ATP (Adenosine Triphosphate) in cancer cells, a Y-type DNA structure specially designed on AuNP and a double-stranded DNA connector are subjected to conformational transformation triggered by miRNA-106a, miRNA-106a is released and circularly amplified, and SERS (Surface Enhanced Raman Scattering) signals are initiated to remarkably enhance the recognition of the cancer cells; meanwhile, DNAzyme generated and amplified through conformation conversion can catalytically crack Survivin mRNA and c-Jun mRNA, and effective double gene silencing treatment is achieved.

Description

technical field [0001] The invention belongs to the technical field of functional nano-probes, and in particular relates to an integrated nano-reagent for cancer diagnosis and treatment and a preparation method and application thereof. Background technique [0002] The invention of an integrated diagnosis and treatment reagent with both diagnostic and therapeutic functions is an important way to achieve synergistic treatment of cancer with high efficiency and low toxicity, and is of great significance for early accurate diagnosis and effective treatment of cancer. The integrated reagent for realizing image-guided cancer therapy provides a powerful means for precise identification of cancer cells, further precise treatment, and timely monitoring of treatment effects. All-in-one reagents that can autonomously initiate treatment modalities after accurate identification of cancer, the cascade process of detection and treatment, have received great attention this year. However, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6816C12N15/11A61K47/52A61K31/713A61P35/00B82Y5/00B82Y30/00
CPCC12Q1/6886C12Q1/6816A61K47/52A61K31/713A61P35/00B82Y5/00B82Y30/00C12Q2600/158C12Q2600/178C12Q2600/136C12Q2525/207C12Q2563/137C12Q2563/155C12Q2565/632
Inventor 宋春元汪联辉董晨
Owner NANJING UNIV OF POSTS & TELECOMM
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