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Serum/plasma miRNA marker associated with breast cancer and application thereof

A marker, breast cancer technology, applied in the field of genetic engineering and oncology

Inactive Publication Date: 2010-12-22
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there are currently no reports of relatively stable biomarkers for the diagnosis of breast cancer. If abnormally expressed serum / plasma miRNAs can be screened out as biomarkers, and corresponding diagnostic kits can be developed, it will be beneficial to breast cancer in my country. The status quo of diagnosis will be a powerful push

Method used

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  • Serum/plasma miRNA marker associated with breast cancer and application thereof
  • Serum/plasma miRNA marker associated with breast cancer and application thereof
  • Serum/plasma miRNA marker associated with breast cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The collection of embodiment 1 sample and the arrangement of sample data

[0079] The inventor has collected a large amount of peripheral blood samples of breast cancer patients and controls from Jiangsu Cancer Hospital and Changzhou Wujin District since 2006 (the samples used for research are collected at the same period, and the conditions of sampling, sub-packaging and storage are uniform). For the sorting of sample data, the inventor selected 196 samples that met the following criteria as experimental samples for Solexa sequencing, TLDA chip detection and a series of subsequent qRT-PCR verifications:

[0080] 1. New breast cancer cases

[0081] 2. No surgery, radiotherapy and chemotherapy before blood collection, no preoperative radiotherapy and chemotherapy

[0082] 3. Healthy female controls matched with the age of the case

[0083] The demographic and clinical data of these samples were collected systematically.

Embodiment 2

[0084] Example 2 Solexa sequencing experiment of miRNA in serum / plasma

[0085] Among the 48 eligible breast cancer patients and 48 healthy female controls described above, the two groups were age-matched. The two groups of people were subjected to Solexa sequencing test to obtain relevant results. The specific steps are:

[0086] 1. Take 50ml of serum from patients in the "breast cancer case" group and the "healthy female control" group, and add an equal volume of Trizol reagent;

[0087] 2. Phase separation: place at room temperature for 15 minutes, then add chloroform according to the volume ratio of 0.2ml chloroform / 1ml Trizol reagent, shake for 15s, room temperature for 15 minutes, centrifuge at 12,000g, 4°C for 15 minutes;

[0088] 3. Transfer the aqueous phase to a new 50ml centrifuge tube, and remove the protein phase in 3 steps of phenol / chloroform;

[0089] 4. RNA precipitation: transfer the aqueous phase to a new centrifuge tube, add isopropanol according to the ...

Embodiment 3

[0097] TLDA chip detection of miRNA in embodiment 3 serum / plasma

[0098] The above 48 breast cancer patients and 48 healthy female controls subjected to Solexa sequencing were detected by TLDA chip to obtain relevant results. The specific steps are:

[0099] 1. Take 600 μl of serum from the "breast cancer case" group and the "healthy female control" group, and add 3 times the volume of Trizol reagent;

[0100] 2. Phase separation: place at room temperature for 15 minutes, add final concentration of 10 -4 Pmol / μl of cel-39 (TAKARA) was used as an internal reference, then chloroform equal to the volume of plasma was added, shaken for 50 s, room temperature for 15 min, 14,000 rpm, 4°C, and centrifuged for 15 min;

[0101] 3. RNA precipitation: transfer the water phase to a new 15ml centrifuge tube, add 1.5 times the volume of the water phase in absolute ethanol, and mix well;

[0102] 4. Enrich RNA with QIAGEN miRNeasy kit: pipette 700 μl of sample into the spin column each t...

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Abstract

The invention belongs to the fields of gene engineering and oncology and discloses a serum / plasma miRNA marker associated with breast cancer and application thereof. The marker is a combination of miR-16, miR-25, miR-222 and miR-324-3p. The maker and primers thereof can be used for preparing a diagnosis kit and assisting early diagnosis of the breast cancer.

Description

field of invention [0001] The invention belongs to the fields of genetic engineering and oncology, and relates to a serum / plasma miRNA marker related to breast cancer and an application thereof. Background technique [0002] Breast cancer is the most common malignancy in women. It is estimated that there were 1.15 million new cases of breast cancer worldwide in 2002, accounting for 23% of all malignant tumors in women, and it is expected to reach 1.4-1.5 million in 2010. In my country, the incidence of breast cancer is relatively low compared with western countries, but it has shown a significant growth trend in the past two decades, with an average annual growth rate of 3-4%, far higher than the world average growth rate of 0.5%. Although breast cancer has a better prognosis, it is still the leading cause of cancer death in women due to its high incidence. In my country's Beijing, Shanghai, Tianjin and other cities, the incidence of breast cancer ranks first and second am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 沈洪兵胡志斌张辰宇马红霞董静
Owner NANJING MEDICAL UNIV
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