Biomarker for breast cancer metastasis and prognosis diagnosis

A biomarker, breast cancer technology, applied in the field of biomedicine

Active Publication Date: 2020-02-25
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on whether the expression of meR342-EZH2 has significant changes in clinical tumors, and whether meR342-EZH2 plays an important role in the progression and metastasis of breast cancer patients

Method used

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  • Biomarker for breast cancer metastasis and prognosis diagnosis
  • Biomarker for breast cancer metastasis and prognosis diagnosis
  • Biomarker for breast cancer metastasis and prognosis diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of antibodies for detecting meR342-EZH2

[0050] The antibody used to detect meR342-EZH2, the amino acid sequence of the antibody is shown in SEQ ID No.1 (TAERIKTPPKRPGGC).

[0051] A method for preparing an antibody for detecting meR342-EZH2, comprising the steps of:

[0052] (1) Using polypeptide synthesis technology to synthesize a polypeptide with an amino acid sequence such as the antibody shown in SEQ ID No.1, so that the purity of the polypeptide is above 85%, and then cross-linking with hemocyanin KLH to obtain the antigen;

[0053] (2) The antigen is mixed with complete adjuvant (CFA) or incomplete adjuvant (IFA) in equal volumes, injected into the animal body, immunized 8 times, and the whole blood of the animal is taken for purification to obtain the antibody.

[0054] In step (2), the animals are New Zealand white rabbits. Take two adult male New Zealand white rabbits purchased from Weitong Lihua Company (the numbers are RB11081 and RB...

Embodiment 2

[0064] Example 2 The relationship between the expression of meR342-EZH2 and the metastasis of breast cancer

[0065] Get 100 cases of breast cancer tissues (wherein, 10 cases of paracancerous tissues, 50 cases of breast cancer tissues in situ, 40 cases of lymph node metastasis breast cancer tissues), adopt the antibody prepared in Example 1, and pass immunohistochemical experiments (using Zhongshan Jinqiao The IHC kit was completed (Zhongshan Jinqiao PV600)) to detect the expression of meR342-EZH2 in normal breast tissue, primary breast cancer samples and metastatic breast cancer samples.

[0066] The operation of the immunohistochemical experiment is as follows:

[0067] (1) Conventional paraffin sections, dewaxed to water, 3% H 2 o 2 Incubate at room temperature for 10 minutes to eliminate endogenous peroxidase activity, wash with PBS 3 times, 5 minutes / time, then boil with antigen retrieval solution (EDTA / sodium citrate) in a pressure cooker for 5 minutes, naturally cool ...

Embodiment 3

[0077] Example 3 meR342-EZH2 promotes the EMT process of breast cancer cells and the invasion and migration of breast cancer cells

[0078] (1) meR342-EZH2 promotes the EMT process of breast cancer cells

[0079] 1) Lentivirus packaging: Spread 293T cells into a 6cm cell culture plate the day before, so that the cell density before transfection reaches 80%; the ratio of related plasmids and transfection reagents for lentivirus packaging is the core plasmid (shEZH2-3'UTR / Vector / Flag-EZH2-WT / Flag-EZH2-R342K): PAX:PMD2G:PEI=4:3:1:3, the total amount of virus plasmid transfected on a 6cm plate is 8μg; add DMEM serum-free medium to a dorf tube After adding the plasmid and PEI, blow and mix gently, and let stand at room temperature for 15 minutes; 6 hours after transfection, discard the medium and replace it with 3ml DMEM complete medium. Collect the first batch of virus liquid after 48 hours of transfection, and then every 24 hours Collect once, replace with 3ml of fresh DMEM com...

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Abstract

The invention relates to a biomarker for breast cancer metastasis and prognosis diagnosis. The biomarker is meR342-EZH2, and through a series of experiments, meR342-EZH2 can promote the EMT process ofbreast cancer and in-vitro invasion and metastasis and in-vivo far-end metastasis of breast cancer cells, through related analysis of case grading, metastasis, prognosis and the like of breast cancerpatients, the meR342-EZH2 high expression is positively correlated with tumor enlargement, high grading and more far-end metastasis sites of the breast cancer patients, and meR342-EZH2 high expression indicates a low five-year survival rate of the breast cancer patients. Therefore, meR342-EZH2 can be used for preparing or screening the breast cancer metastasis diagnostic reagents and breast cancer prognosis diagnostic reagents, and a new diagnosis and evaluation method is provided for breast cancer metastasis and prognosis.

Description

technical field [0001] The invention relates to a biomarker for breast cancer metastasis and prognosis diagnosis, belonging to the technical field of biomedicine. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, accounting for 7-10% of all kinds of malignant tumors in the whole body, ranking first in the incidence of malignant tumors in women. In China, breast cancer is the cancer with the highest incidence rate among women, and the growth rate of the incidence rate is twice the world average level, and the number of deaths accounts for 9.6% of the world's total. The death of breast cancer patients caused by metastasis accounts for more than 90% of all death cases, and the malignant metastasis of breast cancer has become an important cause of death. Clinical data show that a large number of patients with tumor metastasis have metastasized when their original tumor size is small, but because it is not easy to detect, it cannot ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C07K16/40G01N33/574
CPCC12Q1/6886C07K16/40G01N33/57415G01N33/57484C12Q2600/118C12Q2600/136C12Q2600/158G01N2800/52
Inventor 李中伟王典典白津郑骏年侯平甫李敏乐
Owner XUZHOU MEDICAL UNIV
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