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Sequence of aptamer used for targeting human nasopharyngeal carcinoma cell and application thereof

A nucleic acid aptamer and cancer cell technology, applied in the field of cell biology, can solve the problems of low sensitivity and specificity, and achieve the effects of small molecular weight, good permeability, easy synthesis and labeling

Active Publication Date: 2015-06-17
CENT SOUTH UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serological detection of antibodies against Epstein-Barr virus, such as virus capsid protein antigen immunoglobulin A (VCA / IgA), early antigen immunoglobulin A (EA / IgA) and Epstein-Barr virus nuclear antigen immunoglobulin A (EBNA1 / IgA), etc. Used as a serologic screen for high-risk groups, but has low sensitivity and specificity

Method used

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  • Sequence of aptamer used for targeting human nasopharyngeal carcinoma cell and application thereof
  • Sequence of aptamer used for targeting human nasopharyngeal carcinoma cell and application thereof
  • Sequence of aptamer used for targeting human nasopharyngeal carcinoma cell and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0024] Example 2 Detection of dissociation coefficient of sequence S3 on nasopharyngeal carcinoma cell 5-8F by flow cytometry

[0025] The operation of dissociation constant determination is basically the same as that in Example 1. Sequence S3 with different concentrations is arranged in parallel, the fluorescence value of the flow cytometer is used as the ordinate, and the concentration of S3 as the abscissa, Y=Bmax×X / (Kd+X) equation simulates the curve to obtain the dissociation constant of sequence S3. The result is as image 3 As shown, the binding ability of sequence S3 to target cell 5-8F is very strong, and the dissociation constant is 11.9329±1.3997nM.

Embodiment 3

[0026] Example 3 Combination of Detection Sequence S3 and Nasopharyngeal Carcinoma Tissue Section

[0027] The paraffin-embedded nasopharyngeal carcinoma tissue slices were placed in an oven at 60°C for 2 h, and the slices were dewaxed by immersing in xylene twice, 15 min each time; then immersed in absolute ethanol twice, 5 min each time; the slices were sequentially passed through gradient ethanol ( 95%, 90%, 80%, and 70% ethanol once each, 2 min each time), and finally immersed in PBS; the microwave repair method was used to repair the antigen on the slice, and after cooling to room temperature, wash twice with PBS; Block with bovine serum and salmon sperm DNA binding buffer at room temperature for 1 h; remove the blocking solution, add biotin-labeled sequence S3 (200 nM) and incubate at 4°C for 1 h; wash with washing buffer for 3 times, each time for 5 min; then add streptavidin Incubate the quantum dots modified with prime at room temperature for 0.5 h; after washing, cov...

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Abstract

The invention belongs to the technical field of cell biology, and more specifically relates to a sequence of an aptamer used for targeting human nasopharyngeal carcinoma cell and an application thereof. According to the invention, the aptamer capable of specifically combining with human nasopharyngeal carcinoma cells strain 5-8F is screened, so that a sequence S3 capable of specifically combining with 5-8F is obtained. The aptamer of nasopharyngeal carcinoma cell 5-8F has unique stem-loop structure under condition of 0 DEG C, 157mol / L Na<+>, and 0.005mol / L Mg<2+>. The sequence of the aptamer of nasopharyngeal carcinoma cell 5-8F has application prospect on preparation of a nasopharyngeal carcinoma diagnostic reagent, a nasopharyngeal carcinoma treatment medicine, and a nasopharyngeal carcinoma tumour marker, and research of variation of nasopharyngeal carcinoma and normal tissue, nasopharyngeal carcinoma tissue sectioned imaging, nasopharyngeal carcinoma-related in-vivo imaging and nasopharyngeal carcinoma targeting treatment.

Description

technical field [0001] The invention belongs to the technical field of cell biology. It specifically relates to the sequence and application of a nucleic acid aptamer for nasopharyngeal carcinoma cells. Background technique [0002] Nasopharyngeal carcinoma (NPC) is a malignant tumor of epithelial origin with high incidence in southern my country and Southeast Asian countries or regions, with obvious ethnic aggregation and regional characteristics. Nasopharyngeal carcinoma is highly sensitive to radiotherapy and chemotherapy, but its efficacy is related to the degree of disease progression. Unfortunately, most patients with nasopharyngeal carcinoma are at stage III or IV. At this time, the 5-year survival rate with radiotherapy alone is only 35-52%. Combined chemotherapy can effectively improve the therapeutic effect, but the side effects are significantly enhanced. Therefore, the development of early diagnosis methods is critical to improve the overall efficacy. Routin...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N15/14G01N33/569A61K47/26A61P35/00
Inventor 任彩萍贾文婷刘卫东陈玉祥王磊
Owner CENT SOUTH UNIV
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