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Invention and application of PTEN Nedd8 modification as new marker for breast cancer and specific antibody of modification

A breast cancer-specific technology, applied in the field of tumor molecular biology, can solve problems such as difficult to achieve breast-conserving surgery, affecting patients' body and mind and quality of life, and inability to completely remove tumor tissue

Active Publication Date: 2019-11-05
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgical resection of breast cancer often fails to completely remove the tumor tissue due to tumor metastasis, making clinical treatment more difficult, and often relapses after surgery, and it is difficult to achieve breast-conserving surgery in the middle and late stages, which seriously affects the patient's body and mind and quality of life

Method used

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  • Invention and application of PTEN Nedd8 modification as new marker for breast cancer and specific antibody of modification
  • Invention and application of PTEN Nedd8 modification as new marker for breast cancer and specific antibody of modification
  • Invention and application of PTEN Nedd8 modification as new marker for breast cancer and specific antibody of modification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Design and preparation of Nedd8 modified antibody targeting PTEN protein K402 site

[0039] 1. Materials and methods

[0040] 1. Materials

[0041] Normal saline, Freund's complete adjuvant, Freund's incomplete adjuvant, two healthy 6-week-old New Zealand white rabbits (about 2Kg), protein polypeptide,

[0042] 2. Method

[0043] (1) According to the covalent binding of Nedd8 to the K402 site of PTEN protein, the peptide is designed and synthesized. The peptide sequence is CEDQHTQIT-(LALRGG)K-V (SEQ ID No.2, and the peptide sequence in brackets is the 10th lysine Modifier group sequence), this peptide targets the sequence of covalently binding Nedd8 at the K402 site of PTEN protein. This peptide is used as an immunogen for preparing antibodies and an antigen for detecting antibody titers.

[0044] (2) Select two healthy 6-week-old New Zealand white rabbits to adapt to the new living environment, and take blood for the first time after a few days of stabil...

Embodiment 2

[0072] Example 2. Analysis of the changes in the modification degree of Neddylation at the PTEN K402 site in breast cancer tissues and paracancerous tissues

[0073] 1. Materials and methods

[0074] 1. Materials

[0075] Human breast cancer tumor chip (article number: HBreD139Su01 and HBre-Duc090Sur-01, Shanghai Xinchao Biological Co., Ltd., containing clinical information such as patient classification, stage and prognosis), the antibody modified with Nedd8 at K402 site of PTEN protein prepared in Example 1.

[0076] 2. Method

[0077] (1) Put the slices into: bake in a 60°C oven for 20 minutes to melt the wax;

[0078](2) Dewaxing to water: xylene 1→xylene 2→absolute ethanol 1→absolute ethanol 2→95% ethanol→90% ethanol→80% ethanol→70% ethanol→water, each for 5 minutes;

[0079] (3) Wash with PBS for 5 minutes;

[0080] (4) Antigen restoration by microwave: pre-heat the purchased commercial sodium citrate restoration solution (Biyuntian, P0081) in microwave until boiling...

Embodiment 3

[0095] Example 3. Analysis of the changes in the degree of Neddylation modification at the PTEN K402 site in different TNM stages of breast cancer tissues

[0096] 1. Materials and methods

[0097] 1. Materials

[0098] Human breast cancer tumor chip (article number: HBre-Duc170Sur-01 and HBre-Duc090Sur-01, Shanghai Xinchao Biological Co., Ltd., containing clinical information such as patient classification, staging and prognosis), the PTEN protein K402 site Nedd8 prepared in Example 1 Modified antibodies.

[0099] 2. Method

[0100] Immunohistochemistry, the method is the same as in Example 2.

[0101] 2. Results

[0102] Image Pro Plus software counted the IOD (accumulated optical density) level of PTEN protein K402 site Nedd8 modified antibody staining. The analysis results found that the degree of Nedd8 modification at PTEN protein K402 site was positively correlated with TNM stage and pathological grade. Higher, the higher the degree of Nedd8 modification at K402 sit...

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Abstract

Provided is an invention and application of a PTEN Nedd8 modification as a new marker for breast cancer and a specific antibody of the modification. The invention provides application of the Nedd8 modification at the PTEN protein site K402 as the marker in preparing a product used for judging grading and staging of the breast cancer, and application in preparing a product used for predicting breast cancer prognosis. The PTEN Nedd8 modification antibody is further prepared, the PTEN Nedd8 modification antibody can specifically identify the Nedd8 modification at the PTEN site K402, the PTEN Nedd8 modification is enhanced along with increasing of the TNM staging and pathological grading of the tumor malignancy of a breast cancer patient, and the higher the PTEN Nedd8 modification degree is, the poorer the patient's prognosis is. The invention has an important clinical significance for predicting the prognosis conditions of the breast cancer and providing an adjuvant treatment scheme for an individual patient.

Description

technical field [0001] The invention relates to the field of tumor molecular biology, in particular to the invention and application of PTEN Nedd8 modification as a novel marker of breast cancer and its specific antibody. Background technique [0002] Malignant tumor is one of the important diseases that endanger the health and life of our people. In the past 20 years, the incidence of cancer in our country has been increasing year by year. Among them, the incidence rate of lung cancer still tops the list, followed by gastric cancer, colorectal cancer, liver cancer and esophageal cancer. The top ten malignant tumors accounted for 76.39% of the total. Among them, the incidence of female breast cancer is the most, and it shows a younger trend. Breast cancer It has seriously threatened the health of women in our country. PTEN is another tumor suppressor gene that is widely related to tumorigenesis after p53. The embryos of mice with homozygous deletion of PTEN gene are lethal,...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574G01N33/531
CPCG01N33/6893G01N33/6854G01N33/57415G01N33/57484G01N33/531
Inventor 张令强谢萍彭志强陈玉娇杜梦鸽
Owner ACADEMY OF MILITARY MEDICAL SCI
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