Nucleic acid fragment and purpose thereof

A technology of nucleic acid fragments and uses, applied in the field of nucleic acid fragments and their uses, to achieve the effects of low cost, inhibition of proliferation, and large molecular weight

Active Publication Date: 2014-10-22
成都杰芈卡科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, severe resistance to tamoxifen necessitates the development of new antiestrogens

Method used

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  • Nucleic acid fragment and purpose thereof
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  • Nucleic acid fragment and purpose thereof

Examples

Experimental program
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Embodiment 1

[0030] Example 1 Preparation of nucleic acid fragment (ER-aptamer) of the present invention and identification of its binding ability to estrogen receptor

[0031] 1. Preparation of nucleic acid aptamers for estrogen receptors of the present invention

[0032] Using the in vitro screening technique SELEX (System Evolution of Ligands by Exponential Enrichment) to screen the nucleic acid aptamer of the estrogen receptor, a nucleic acid fragment with high affinity to the estrogen receptor was obtained, and its sequence (SEQ ID NO.1) is as follows:

[0033] 5'-GTCAGGTCACAGTGACCTGATCA AAGTTAATG-3'.

[0034] Sent to Shenggong Bioengineering (Shanghai) Co., Ltd. for synthesis.

[0035] 2. Verification of the binding ability of nucleic acid aptamers to estrogen receptors

[0036] In order to determine the affinity of the nucleic acid aptamer shown in SEQ ID NO.1 above to the estrogen receptor, an ELISA-based ligand-receptor binding assay method is used for determination.

[0037] E...

Embodiment 2

[0046] Embodiment 2 Detection kit of the present invention

[0047] 1. Kit 1

[0048] Kit composition (50 servings):

[0049]

[0050] How to use the kit:

[0051] (1) Prepare slices of tissue samples to be seen;

[0052] (2) Prepare biotin-labeled ER-nucleic acid aptamer with DNA-protein binding buffer, and incubate overnight at 4°C with the slice prepared in step (1). Then wash with 0.01M phosphate buffered saline (PBS);

[0053] (3) Dilute the Streptavidin-horseradish peroxidase (HRP) conjugate (Sigma Chemical Co, St.Louis, Mo, USA) with PBS buffer to 1:1000, and combine with the biotin-labeled ER-Aptamer Reaction. Color was developed with DAB peroxidase substrate solution (Dako, Denmark A / S).

[0054] (4) Observation under an optical microscope.

[0055] 2. Kit 2

[0056] Kit composition (50 servings):

[0057]

[0058] How to use the kit:

[0059] (1) Prepare slices of tissue samples to be seen;

[0060] (2) Configure HRP-labeled ER-nucleic acid aptamer w...

experiment example 1

[0072] Experimental Example 1 Detection experiment of breast cancer and endometrial cancer

[0073] ER-nucleic acid aptamer: the nucleic acid fragment prepared in Example 1 of the present invention.

[0074] 1. Experimental steps:

[0075] (1) Section preparation: fix the sections of 4 μm thick paraffin-embedded tissue blocks on polylysine-coated glass slides; dewax with xylene and hydrate with graded ethanol; Antigen retrieval treatment was performed at 120°C (autoclave) in buffer (pH6.0) for 5 minutes; endogenous peroxidase activity was blocked with 0.03% hydrogen peroxide solution containing sodium azide, and placed at room temperature for 30 minute.

[0076] (2) Prepare the biotin-labeled ER-nucleic acid aptamer with DNA-protein binding buffer (0.01MTris-hydrochloric acid, pH7.5, 1mM magnesium chloride, 0.5mM DTT, 0.5mM EDTA and 50nMNaCl) (1: 50), that is, take 10 μl of biotin-labeled ER-nucleic acid aptamer and add it to 490 μl of DNA-protein binding buffer, and mix we...

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Abstract

The invention discloses a nucleic acid fragment, a nucleotide sequence of the nucleic acid fragment is shown as a SEQ ID NO.1. The invention also discloses a purpose of the nucleic acid fragment in preparation of a reagent for diagnosing breast cancer and / or endometrial carcinoma, and a kit composed of the diagnostic reagent. The invention also discloses a purpose of the nucleic acid fragment in preparation of an estrogen receptor antagonist, the nucleic acid fragment can be used for preparing a medicine for treating breast cancer, and has practical application value.

Description

technical field [0001] The present invention relates to a nucleic acid fragment and its application. Background technique [0002] Since the late 1970s, the incidence of breast cancer has been ranked first in female tumors worldwide. Although my country is not a country with a high incidence of breast cancer, the incidence rate has been increasing year by year. According to the expression of estrogen receptor (ER), breast cancer can be divided into estrogen receptor-dependent breast cancer and non-hormone receptor-dependent breast cancer. Estrogen receptor-dependent breast cancer expresses estrogen receptors and depends on estrogen for growth. It is sensitive to hormones and weakly resistant, and can be treated with anti-estrogen drugs, while non-hormone receptor-dependent breast cancer does not express estrogen receptors The body does not rely on estrogen for growth, has poor sensitivity and strong resistance to hormones, and cannot be treated with anti-estrogen drugs. Ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 叶尚勉
Owner 成都杰芈卡科技有限公司
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