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Aptamer and application in preparing medicine or product for treating leukaemia thereof

A nucleic acid aptamer and leukemia technology, applied in the fields of biomedicine and clinical medicine, to achieve high-affinity effects

Active Publication Date: 2013-11-27
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The therapeutic effect of nucleic acid aptamer TY04 on leukemia has not been reported yet, which will provide a new strategy for the treatment of leukemia

Method used

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  • Aptamer and application in preparing medicine or product for treating leukaemia thereof
  • Aptamer and application in preparing medicine or product for treating leukaemia thereof
  • Aptamer and application in preparing medicine or product for treating leukaemia thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] In order to clarify whether the nucleic acid aptamer TY04 has an effect on the proliferation of lymphocytic leukemia cell lines, we used the CCK-8 method to detect the 4μM TY04 treatment of various leukemia cell lines (K562, HL-60, Jurkat, CEM, ARH77 ) and normal human immortalized B lymphocytes (the concentration of the tested cells was 1×10 5 Individuals / ml) The relative survival rate after 96h. After three independent repeated experiments, it was found that TY04 treatment can significantly inhibit the proliferation of various leukemia cell lines (see figure 1 Generally speaking, if the IC50 value at the cell level is within 10 μM, it has anti-tumor development potential). CCK-8 was further used to detect different concentrations of nucleic acid aptamer TY04 (0, 0.5, 1.0, 2.0, 4.0 μM) to treat lymphocytic leukemia ARH77 cells and normal human immortalized B lymphocytes (the concentration of the tested cells was 1× 10 5 per ml) after 96 hours, after three independen...

Embodiment 2

[0022] In order to clarify the binding of nucleic acid aptamer TY04 to lymphocytic leukemia ARH77 cells, the nucleic acid aptamer TY04 with FITC fluorescent labeling was mixed with ARH77 cells (test cells) at different concentrations (0, 25, 125, 250, 500 nM) The concentration is 1×10 5 cells / ml) after incubation, the binding of TY04 to the surface of ARH77 cells was detected by flow cytometry. The result is as image 3 As shown, the fluorescent nucleic acid aptamer TY04 has significant binding to ARH77 cells, and the binding strength increases with the increase of the concentration.

Embodiment 3

[0024] In order to observe the binding site between TY04 and ARH77 cells, 250nM FITC fluorescently labeled nucleic acid aptamer TY04 was incubated with ARH77 cells at 4°C and 37°C respectively, and observed under an upright fluorescence microscope after preparation. The results are as follows: Figure 4 , A is a white light picture, and B is a fluorescent picture. It can be seen that the nucleic acid aptamer TY04 is mainly distributed on the ARH77 cell membrane. It is speculated that it may bind to membrane proteins through its unique three-dimensional conformation, interfere with the functional activity of membrane proteins, and disrupt the signal transduction in cells. Conducting pathway, and ultimately produce the biological effect of inhibiting the proliferation of ARH77 cells.

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Abstract

The invention provides an aptamer and application in preparing medicine or product for treating leukaemia thereof. The aptamer is TY04 and is a section of ssDNA, containing 83 bases. The action principle of the aptamer is that the aptamer can present cell specificity and cell sequence specificity and is combined with a leukaemia cell, blocks a cell period process in a G2 / M stage via elevating the expression of protein p21 relative to the cell periodic duty of a leukaemia cell ARH77, thereby restraining the proliferation of the leukaemia cell. So far, news that the aptamer is applied to the treatment of the leukaemia is not reported at home and abroad. The discovery and the application of the aptamer provide a new strategy for treating the leukaemia in the future.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and clinical medicine, and relates to a nucleic acid aptamer related to the treatment of leukemia and its application in the preparation of medicines or products for treating leukemia. Background technique [0002] Nucleic acid aptamer (aptamer) is a short single-stranded DNA or RNA molecule with recognition function screened by an artificial screening technique that simulates the natural evolution process-exponential enrichment ligand system evolution (SELEX), and can form a specific DNA or RNA molecule by folding. The three-dimensional structure binds to the target molecule. It has the characteristics of high affinity, high specificity, non-immunogenicity, easy synthesis, transformation and labeling, good biochemical stability, reversible denaturation and renaturation, and can be amplified and cut by enzymes, especially some nucleic acids Aptamers can specifically bind to their target prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115A61K48/00A61P35/02
Inventor 谭蔚泓叶茂刘静宋健辉戴红娟
Owner HUNAN UNIV
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