Applications of eukaryotic translation elongation factors in breast cancer detection reagent

A translation elongation factor and breast cancer technology, which is applied in the application field of eukaryotic translation elongation factor in the detection of breast cancer reagents, and achieves the effect of convenient and easy diagnosis.

Inactive Publication Date: 2017-06-20
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is no report on the application of SNP in the diagnosis of breast cancer. If the SNP of breast cancer susceptibility can be screened out as a biomarker, and a corresponding diagnostic kit can be developed, it will greatly promote the current situation of early diagnosis of breast cancer in my country. , and open up new ways for its drug screening, efficacy evaluation and targeted therapy

Method used

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  • Applications of eukaryotic translation elongation factors in breast cancer detection reagent
  • Applications of eukaryotic translation elongation factors in breast cancer detection reagent
  • Applications of eukaryotic translation elongation factors in breast cancer detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The collection of embodiment 1 sample and the arrangement of sample data

[0053] From January 2010 to December 2015, the inventor collected a large number of blood samples from patients with new breast cancer in Shenzhen Second People's Hospital. After sorting out the sample data, the inventor selected 25 samples that met the following criteria At the same time, 10 healthy women aged 25-55 were selected as controls for whole-exome microarray testing. The sample selection criteria were as follows:

[0054] 1. Breast cancer cases diagnosed by pathology, among which 3 patients have a family history of cancer and are marked as X1, X2, and X3 respectively;

[0055] 2. Have not received radiotherapy or chemotherapy before blood collection, and have no previous history of cancer;

[0056] 3. Healthy female controls matched with the age of the case

[0057] The demographic data and clinical data of these samples were collected systematically.

Embodiment 2

[0058] Example 2 Extraction and Purification of Peripheral Blood DNA

[0059] Among the above-mentioned 25 eligible breast cancer patients and 10 healthy female controls, the age balance of the two groups was comparable.

[0060] The specific steps are:

[0061] 1. Add hemolysis reagent (i.e., lysate, 40 parts) to the peripheral blood stored in a 2mL cryopreservation tube. The volume of the solution was adjusted to 2000mL, the same below), and it was completely transferred after inverting to mix well.

[0062] 2. Removal of red blood cells: Fill the 5mL centrifuge tube to 4mL with hemolysis reagent, mix by inverting, centrifuge at 4000rpm for 10 minutes, and discard the supernatant. Add 4 mL of hemolysis reagent to the precipitate, invert and wash again, centrifuge at 4000 rpm for 10 minutes, and discard the supernatant.

[0063] 3. Extract DNA: add 1mL extract solution (each 300mL contains 122.5mL0.2M sodium chloride, 14.4mL0.5M ethylenediaminetetraacetic acid, 15mL10% sod...

Embodiment 3

[0068] Example 3 Whole Exome Detection of SNP

[0069] The two groups of people in Example 2 were detected by the whole exome chip to obtain relevant results.

[0070] 1. Library construction

[0071] Beijing Novogene Technology Co., Ltd. uses Agilent's liquid-phase chip capture system to efficiently enrich human DNA from the entire exon region, and then perform high-throughput and high-depth sequencing on the Illumina Hiseq platform. The Agilent SureSelect Human All ExonV5 kit was used for library construction and capture experiments, the reagents and consumables recommended in the instructions were strictly used, and the operation was performed according to the latest optimized experimental procedures.

[0072] The basic process of the experiment: Genomic DNA was randomly broken into fragments with a length of 180-280bp by a Covaris crusher, and after end repair and A-tailing, adapters were connected to both ends of the fragments to prepare a DNA library. After pooling, th...

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Abstract

The invention discloses applications of eukaryotic translation elongation factors in the breast cancer detection reagent, wherein the eukaryotic translation elongation factors comprise the eukaryotic translation elongation factor 1delta and the eukaryotic translation elongation factor 2. The invention further discloses the reagent for detecting the genotype of the susceptible SNP site of the eukaryotic translation elongation factors, and a kit prepared by adopting the reagent. Through the development and applications of the SNP biomarker of the eukaryotic translation elongation factors and the diagnostic kit, the diagnosis for breast cancer is convenient and feasible, a foundation is laid for clinicians to rapidly know the disease conditions of patients, and for evaluating the clinical treatment effects, and assistance is offered for searching for novel micromolecular drug targets with the potential treatment value.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of eukaryotic translation elongation factor in detecting breast cancer reagents. Background technique [0002] Breast cancer is a systemic disease, and its occurrence and development is a complex process involving multiple factors and links, including the activation of oncogenes and the inactivation of tumor suppressor genes. Therefore, gene mutation plays a very important role in the occurrence and development of breast cancer. [0003] Breast cancer is a multifactorial genetic variation disease, less than 10% is caused by a single gene defect. With the development of high-throughput gene technology, more and more breast cancer-related genes have been discovered, and potential genetic variations (single nucleotide polymorphisms and copy number variations) in these genes may cause differences in the therapeutic effects of breast cancer drugs . Due to the ex...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 付利何劲松谭斌斌陈伟财邹畅向迪刘蓓蕾罗雪莹潘悦
Owner SHENZHEN UNIV
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