Molecular marker for breast carcinoma diagnosis

A breast cancer, thioredoxin technology, applied in the direction of analysis materials, measuring devices, instruments, etc., can solve the problem of breast cancer correlation that has not been seen in thioredoxin

Inactive Publication Date: 2009-07-29
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there is no report on the correlation between thioredo...

Method used

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  • Molecular marker for breast carcinoma diagnosis

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Embodiment 1

[0026] Preparation of protein samples from normal surrounding tissues and mass tissues of breast cancer:

[0027] Breast tissue: The samples were from the First Affiliated Hospital of Kunming Medical College, Yunnan Province. All tumors have been pathologically diagnosed.

[0028] The method for preparing protein samples from normal surrounding tissues and mass tissues of breast cancer is as follows:

[0029] 1. The fresh tissue pieces removed by surgery are stored in a refrigerator at -80℃.

[0030] 2. When preparing protein, cut out tissue pieces of about 5 mm, grind them into cell pellets with a grinder on ice, and add 120 microliters of cell lysate (sodium chloride, NP-40, Tris, PMSF, aprotimin, NaN3 / 100 Ml), placed on ice for 50 minutes, then centrifuged at 12000r / min at 4°C for 15 minutes, and the supernatant was stored in a refrigerator at -80°C.

Embodiment 2

[0032] Western blot verification of thioredoxin and thioredoxin binding protein 2:

[0033] In order to confirm the differential expression of thioredoxin and thioredoxin binding protein 2, samples of the mass tissue and corresponding normal peripheral tissue proteins of 30 breast cancer patients were taken, and anti-thioredoxin and thioredoxin binding protein were used. The antibody of 2 is subjected to western blot analysis, and the specific process is briefly described as follows:

[0034] Take 7 ug total protein load for each sample, separate by 15% SDS-PAGE, transfer to the PVDF membrane (purchased from Millipore), and incubate the PVDF membrane overnight with skim milk at 4°C. Wash three times with TPBS for 5 minutes each time. The primary antibody was a mouse anti-human thioredoxin antibody (provided by Professor Yodoi Yuji, Kyoto University, Japan), incubated for 1 hour at room temperature, and washed three times with TPBS for 5 minutes each. Secondary antibody anti-mouse ...

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Abstract

The invention discloses a molecular marker for diagnosing breast cancer, which is used for detecting expression of the thioredoxin (TRX) and the thioredoxin binding protein 2 (TRX-binding protein-2, TBP-2) in surrounding normal breast tissue and mass tissue. A protein immunoblot experiment is used for detecting the expression amount of the thioredoxin and the thioredoxin binding protein 2; in breast mass tissue, the expression of the thioredoxin is increased and the expression of the thioredoxin binding protein 2 is decreased; as significant difference exists between the breast mass tissue and the normal breast tissue, the thioredoxin and the thioredoxin binding protein 2 can be used as a new molecular marker of breast cancer and used for the clinical diagnosis of breast cancer.

Description

Technical field [0001] The present invention belongs to the field of biotechnology. Specifically, it relates to a Western blotting method for diagnosing breast cancer, and thioredoxin and thioredoxin binding protein 2 are used as protein molecular markers for detecting breast cancer. Clinical application. Background technique [0002] Breast cancer is one of the main diseases that endanger women's health. About one million women worldwide suffer from breast cancer every year. Men can also get breast cancer, but the chance of men getting breast cancer is nearly 100 times less than that of women. The onset of breast cancer is mostly between the ages of 40 and 60. The reason for its occurrence is still inconclusive. It is generally believed that it is related to changes in sex hormones and has a certain familial nature. Early breast cancer may not have any conscious symptoms. In the late stage of the disease, breast lumps may appear. The texture of the lumps is hard and tough, the b...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 白洁党英男蒋爱梅李彦辉
Owner KUNMING UNIV OF SCI & TECH
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