Application of deubiquitinating enzyme USP25 in preparation of biomarker for diagnosing breast cancer

A deubiquitinating enzyme and biomarker technology, applied in the field of deubiquitinating enzyme USP25, to prepare biomarkers for the diagnosis of breast cancer, can solve the problems of poor breast cancer effect, achieve good HR repair ability, improve The effect of repairing ability and enhancing resistance ability

Pending Publication Date: 2022-01-14
SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Aiming at the above-mentioned technical problems in the prior art, the present invention provides the use of the deubiquitinating enzyme USP25 in the preparation of biomarkers for the diagnosis of breast cancer. Cancer's Poor Technical Issues

Method used

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  • Application of deubiquitinating enzyme USP25 in preparation of biomarker for diagnosing breast cancer
  • Application of deubiquitinating enzyme USP25 in preparation of biomarker for diagnosing breast cancer
  • Application of deubiquitinating enzyme USP25 in preparation of biomarker for diagnosing breast cancer

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Experimental program
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Effect test

Embodiment 1

[0023] The amino acid sequence of the protein encoded by USP25 is shown in SEQ ID NO.1, and the gene sequence is shown in SEQ ID NO.2. Abcam (ab187156) monoclonal antibody (purchased from Shanghai Pufei Biotechnology Co., Ltd.) was used to detect USP25.

Embodiment 2

[0024] Example 2 Deubiquitinating enzyme USP25 affects DNA damage repair ability

[0025] The most serious of the types of DNA damage are DNA double-strand breaks (DSBs), which can cause cell death if not repaired in time. The generation of DSB will activate the cell's DNA damage response (DDR) mechanism. DDR senses, amplifies and transduces DNA damage signals by rapidly recruiting a large number of proteins. These recruited proteins form foci of clusters (foci) at DSBs, a hallmark of DSB formation.

[0026] Ionizing radiation is the most common method of inducing DNA double-strand breaks. Using shRNA to knock down USP25 in U2OS cells, Western results showed that U2OS cell lines have been stably depleted of USP25 ( figure 1 A). The control and USP25-knockout cells were irradiated with ions, and immunofluorescence experiments showed that after depletion of USP25, the ability of Rad51 and Rpa32 to form foci was significantly increased, and more were recruited to DNA damage co...

Embodiment 3

[0027] Example 3 USP25 is regulated by DNA damage response

[0028] Phosphorylation is an important type of post-translational modification in the DNA damage response pathway, which can affect the activity and stability of proteins. Therefore, detecting the change of USP25 phosphorylation level can reflect whether USP25 is regulated by DNA damage response. Such as figure 2 As shown, under the induction of ionizing radiation, the USP25 in the cells was immunoprecipitated, and the ATM substrate-specific phosphorylated antibody p-SQ / TQ was used to detect that USP25 could be phosphorylated by ATM ( figure 2 A), but after mutating the phosphorylation sites S85, S258 and T523 of USP25, it was found that after the T523 position of USP25 was mutated, the phosphorylation level was no longer increased ( figure 2 B).

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Abstract

The invention provides an application of deubiquitinating enzyme USP25 in preparation of a biomarker for diagnosing breast cancer. The invention also provides application of the reagent for detecting the deubiquitinating enzyme USP25 in a kit for diagnosing breast cancer. The action principle that a homologous recombination repair mechanism regulates and controls breast cancer drug resistance is utilized, it is identified that the deubiquitinating enzyme USP25 is related to a homologous recombination repair pathway, the cell homologous recombination repair capacity can be remarkably improved by knocking out the USP25, and the resistance capacity of breast cancer cells to a PARP inhibitor is remarkably enhanced. Besides, IHC detection proves that USP25 is highly expressed in breast cancer tissues, and database analysis further shows that a breast cancer patient with simultaneous low expression of USP25 and BRCA1 has better HR repair capacity and is more tolerant to chemotherapeutic drugs. Therefore, the invention finds a novel molecular marker for breast cancer diagnosis and personalized treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to deubiquitinating enzyme USP25, specifically the application of deubiquitinating enzyme USP25 in preparing biomarkers for diagnosing breast cancer. Background technique [0002] In 1990, research discovered a gene directly related to hereditary breast cancer, BRCA1, which is located on chromosome 17 of the human nucleus. If the structure of the BRCA1 gene is mutated, its function of inhibiting tumorigenesis will be affected, which will increase the risk of breast cancer and ovarian cancer. Studies have shown that for those with BRCA1 gene mutation, the risks of breast cancer and ovarian cancer are 50%-85% and 15%-45%, respectively. In the general population, about 12.0% of women may develop breast cancer. BRCA1 gene mutation makes 4-7 times increased risk of breast cancer. In terms of ovarian cancer, only 1.4% of ordinary women have the risk of ovarian cancer, and the BRCA1 gene mutatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574G01N33/573
CPCC12Q1/6886G01N33/57415G01N33/57484G01N33/573C12Q2600/158C12Q2600/156C12Q2600/106C12Q2600/118G01N2333/916G01N2333/948G01N2333/9015G01N2333/9108
Inventor 李昀辉袁健李磊王新舒陈玉平
Owner SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE
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