Application of gene marker in diagnosis and treatment of breast cancer

A breast cancer, gene technology, applied in the field of biomedicine, can solve the problems of breast cancer metastasis and recurrence that cannot be cured

Active Publication Date: 2018-04-06
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although early detection, early treatment, early surgery and other strategies have increased the five-year...

Method used

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  • Application of gene marker in diagnosis and treatment of breast cancer
  • Application of gene marker in diagnosis and treatment of breast cancer
  • Application of gene marker in diagnosis and treatment of breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Screening for gene markers associated with breast cancer

[0078] 1. Sample collection

[0079] The paracancerous tissue and breast cancer tissue samples of 8 breast cancer patients with definite pathological diagnosis were collected, and the sample name, tissue type, number, sampling date, sample processing process, etc. were written, and the patients signed the informed consent. All were approved by the organizational ethics committee.

[0080] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0081] Take out the tissue samples frozen in liquid nitrogen, put the tissue samples into a pre-cooled mortar for grinding, and wait until the tissue samples are powdered:

[0082] 1) Add Trizol and place at room temperature for 5 minutes;

[0083] 2) Add 0.2ml of chloroform, vibrate the centrifuge tube vigorously, mix well, and place it at room temperature for 5-10min;

[0084] 3) Centrifuge at 12000rpm for 15min, transfer the upper...

Embodiment 2

[0096] Example 2 QPCR sequencing to verify the differential expression of the LINC02544 gene

[0097] 1. Large-scale QPCR verification of the differential expression of the LINC02544 gene. According to the sample collection method in Example 1, 50 cases of breast cancer paracancerous tissues and 50 cases of breast cancer tissues were selected.

[0098] 2. The RNA extraction steps are as described in Example 1.

[0099] 3. QPCR

[0100] 1) Reaction system:

[0101] Add 1 μl of RNA template, 1 μl of random primers, add double distilled water to 12 μl, mix well, centrifuge at low speed at 65°C for 5 min, then place on ice to cool.

[0102] Continue to add the following components to the 12 μl reaction solution:

[0103] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and centrifuge;

[0104] 2) Reverse transcription reaction conditions

[0105] 5 minutes at 25°C, 60 minutes at 42°C, and 5 minutes...

Embodiment 3

[0124] Example 3 Expression of LINC02544 in breast cancer cell lines

[0125] 1. Cell culture

[0126] Cultivate human breast cancer cell lines MCF-7, SK-BR-3, MDA-MB-231 and breast normal epithelial cell line MCF-10A, wherein MDA-MB-231 is cultured in L15 medium with 10% fetal bovine serum , SKBR3 was cultured in RPMI-1640 medium containing 10% fetal bovine serum, and MCF-7 and normal mammary epithelial cell line MCF-10A were cultured in DMEM medium containing 10% fetal bovine serum. Add 1% P / S to the culture medium. At 37°C, 5% CO 2, Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0127] 2. RNA extraction and concentration determination

[0128] 1) Wash the cells to be collected with PBS, add an appropriate amount of RNAiso Plus, incubate on ice for 2 minutes to lyse, scrape off the cells at the bottom of the bottle, pipette repeatedly with a mic...

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Abstract

The invention discloses an application of a gene marker in diagnosis and treatment of breast cancer. The gene is LINC02544, the invention firstly discovers that expression of LINC02544 in a breast cancer patient is obviously up-regulated, and verification is further carried out through QPCR, so that LINC02544 can be applied to diagnosis of the breast cancer as a biomarker. The invention also discloses that proliferation of breast cancer cells can be effectively inhibited and migration and attack rates of the breast cancer cells can be reduced by reducing expression of LINC02544, so that LINC02544 can be applied to treatment of the breast cancer and shift thereof as a drug target.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of gene markers in breast self-diagnosis and treatment, in particular to the application of LINC02544 in breast cancer diagnosis and treatment. Background technique [0002] Breast cancer is the most common malignant tumor in women worldwide, accounting for about 22% of all malignant tumors in women, and is one of the most important diseases that endanger women's health. In the past, the incidence of breast cancer in China and other developing countries was low. However, with changes in society, economy, environment and many other factors, the incidence of breast cancer has increased rapidly around the world. In just 10 years, the overall incidence of breast cancer has increased. Breast cancer in China is growing rapidly at an annual rate of 2.7%. Epidemiological data show that abnormal amplification and mutation of genes and genetic susceptibility changes also play an im...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/136C12Q2600/158
Inventor 石小峰孙锦云
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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