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Application of nucleic acid aptamer in recognizing and being combined with bladder cancer cells

A nucleic acid aptamer and cell technology, applied in biochemical equipment and methods, material testing products, biological tests, etc., can solve the problems of inconvenience and poor specificity in identifying and binding bladder cancer cells

Inactive Publication Date: 2019-09-24
谷雨中科(北京)医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For this reason, the embodiment of the present invention provides a nucleic acid aptamer and its application in identifying and binding bladder cancer cells, so as to solve the problems of inconvenient detection and binding of bladder cancer cells in vitro and poor specificity in the prior art

Method used

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  • Application of nucleic acid aptamer in recognizing and being combined with bladder cancer cells
  • Application of nucleic acid aptamer in recognizing and being combined with bladder cancer cells
  • Application of nucleic acid aptamer in recognizing and being combined with bladder cancer cells

Examples

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Embodiment 1

[0032] Embodiment 1, the screening method of bladder cancer cell-specific nucleic acid aptamer

[0033] 1. Establishment of single-stranded DNA library and amplification primers by chemical synthesis in vitro

[0034] The sequence of the single-stranded DNA library is: 5'GGTAGACATGACTAATCGGTCTA-(25N)-TTCTGCGACAGTCTGACTACACGG-3';

[0035] The nucleotide sequence of the upstream primer is shown in SEQ ID NO.2; the nucleotide sequence of the downstream primer is shown in SEQ ID NO.3.

[0036] 2. Using cell-SELEX technology to screen bladder cancer cell-specific nucleic acid aptamers

[0037] A synthetic single-stranded DNA library was used, EJ cells were used as positive screening cells, and the normal human bladder cell line HCV29 was used as negative screening cells. Cell-SELEX technology was used to screen a bladder cancer cell-specific nucleic acid aptamer. The specific steps are:

[0038] 2.1 Take the 1000pmol single-stranded DNA (ssDNA) library synthesized in vitro, cent...

Embodiment 2

[0049] Example 2. Analysis of the Expression of Nucleic Acid Aptamers in Human Bladder Cancer Tissue and Normal Bladder Tissue

[0050] 1. Materials and methods

[0051] 1. Materials: Tissue samples come from inpatient surgical resection samples of bladder cancer patients, and each pair contains bladder cancer tissue and paired normal bladder tissue.

[0052] 2, method: (1) get the tumor tissue of bladder cancer patient and normal person to make pathological section in normal tissue; (2) get bladder cancer tissue as experimental group, normal bladder tissue as contrast, screen with embodiment 1 of the present invention The specific bladder cancer nucleic acid aptamer was used as the primary antibody for immunohistochemical staining, and the specific steps were as follows:

[0053] 2.1. The immunohistochemical pretreatment method was used to process the pathological sections of the experimental group and the control group.

[0054] 2.2. Add the specific bladder cancer nucleic...

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Abstract

The embodiment of the invention discloses a nucleic acid aptamer. The nucleotide sequence of the nucleic acid aptamer is shown in SEQ ID NO.1. By synthesizing a random single-stranded DNA library and a primer, an EJ cell, namely a human bladder cancer cell line is taken as a positive selection cell, and an HCV29 cell, namely a human bladder normal cell line is taken as a reverse selection cell. Firstly, positive selection is performed, the obtained single-stranded DNA library is subjected to negative control cell screening to remove non-specific single-stranded DNA, and cell-SELEX circulation is performed multiple times, and the finally obtained specific single-stranded DNA of the bladder cancer cell line and bladder cancer tissue cells are subjected to positive binding screening to obtain a precise specific nucleic acid aptamer library binding to EJ cells.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of biological detection, in particular to the application of a nucleic acid aptamer in identifying and binding to bladder cancer cells. Background technique [0002] Bladder cancer is the most common malignant tumor in the urinary system, which is multifocal and prone to recurrence after surgery. Cystoscopy and urine exfoliation cytology are the gold standard for the diagnosis of bladder cancer, but the invasiveness of cystoscopy and the low sensitivity and specificity of urine exfoliation cytology make it difficult to detect bladder cancer early and postoperative recurrence lesion. The detection of early diagnosis or postoperative recurrence of non-basal invasive bladder cancer usually requires cystoscopy, X-ray, CT, ultrasound and other methods. It is an invasive examination, and patients have a certain resistance to it, which leads to a decline in medical compliance and makes p...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N33/53
CPCC12N15/115C12N2310/16G01N33/53
Inventor 李翀
Owner 谷雨中科(北京)医疗科技有限公司
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