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Screening and application of nucleic acid aptamer specifically targeting breast cancer cell line MCF-7

A nucleic acid aptamer, MCF-7 technology, applied in the field of biomedicine, can solve the problems of long screening time, low screening efficiency, complicated process, etc., and achieve the effects of simple and convenient operation, simple structure and short cycle.

Active Publication Date: 2019-08-30
CHANGCHUN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] SELEX technology is a general and powerful tool for studying aptamers, but the traditional SELEX technology has low screening efficiency, long screening time and complicated process

Method used

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  • Screening and application of nucleic acid aptamer specifically targeting breast cancer cell line MCF-7
  • Screening and application of nucleic acid aptamer specifically targeting breast cancer cell line MCF-7
  • Screening and application of nucleic acid aptamer specifically targeting breast cancer cell line MCF-7

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Embodiment Construction

[0023] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0024] see Figure 1~4 , in the embodiment of the present invention, a nucleic acid aptamer screening specifically targeting breast cancer cell line MCF-7, using cell-SELEX technology, using human breast cancer cell line MCF-7 as target cells, normal human breast cells MCF-10A was used as a control cell for breast cancer cell nucleic acid aptamer screening, and the specific steps were as follows:

[0025] (1) MCF-7 cells were used for the first three rounds o...

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Abstract

The invention discloses screening and application of a nucleic acid aptamer specifically targeting a breast cancer cell line MCF-7. The nucleotide sequence of the nucleic acid aptamer refers to a DNAfragment represented by the following sequence: a sequence 9-A: 5'-AGGAGCACGACTCTGACGTAGGATCGAGACGAGGTACGTAT-3'. The screening employs a cell-SELEX technology and comprises the following steps: firstsynthesizing an initial library; using the breast cancer cell line MCF-7 as target cells, and conducting incubation with the library; and conducting positive screening, elution, PCR amplification andsingle-stranded DNA preparation. For breast cancer cell detection, the nucleic acid aptamer has the advantages of simple and convenient operation, low cost, short cycle, and high accuracy; and targetmolecules combined with the nucleic acid aptamer of the invention are used for research, and tumor markers can be obtained. An effective tool and means are provided for early diagnosis of breast cancer, breast cancer typing and other applications, and the nucleic acid aptamer can be used for early diagnosis of breast cancer, preparation of breast cancer diagnostic drugs and kits.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to screening of a nucleic acid aptamer specifically targeting breast cancer cell line MCF-7 and its application. Background technique [0002] Nucleic acid aptamer (aptamer), also known as aptamer, aptamer, etc., is a type of small molecule RNA or single-stranded DNA, usually composed of 20-100 bases, from a synthetic RNA / DNA library Screen and isolate single-stranded oligonucleotides that can bind to target substances with high specificity and high sensitivity. Although the binding method of aptamers is similar to that of antibodies, the unique properties of nucleic acid aptamers surpass those of antibodies in many aspects, and are called "chemical antibodies" by scholars. Compared with antibodies, nucleic acid aptamers are completely screened in vitro, and have obvious advantages in terms of production cost, stability, ease of modification, and tissue penetration. Nucleic ac...

Claims

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Application Information

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IPC IPC(8): C12N15/115C12N15/10G01N33/574
CPCC12N15/111C12N15/115C12N2310/16C12N2320/13G01N33/57415
Inventor 金丽虹申炳俊张佳佳李梦媛逄博刘莹
Owner CHANGCHUN UNIV OF SCI & TECH