Method for researching differentially expressed proteins of triple negative breast cancer exosomes by using iTRAQ technology

Abstract

The invention discloses a method for researching differentially expressed proteins of triple negative breast cancer exosomes by using an iTRAQ technology. The method comprises the following steps of (1) extracting and treating exosomes and exosome proteins; (2) carrying out enzymatic hydrolysis and desalination on exosome protein samples obtained in the step (1) to obtain polypeptides; (3) carrying out iTRAQ labelling and mixing on the polypeptides obtained in the step (2); (4) carrying out HPLC separation and LC-MS analysis on a polypeptide-peptide mixture labelled in the step (3); and (5) carrying out bioinformatics analysis and quantitative analysis on the obtained mass spectrometry identification result to obtain the differentially expressed proteins of the triple negative breast cancer exosomes. Research on triple negative breast cancer exosome proteome can be quickly and effectively carried out by combining the iTRAQ technology disclosed by the invention with an MS / MS technology,and a reference is provided for researching a disease mechanism of a triple negative breast cancer and discovering triple negative breast cancer-related biomarkers.

Description

technical field [0001] The invention relates to the field of triple-negative breast cancer, in particular to a method for studying differentially expressed proteins in exosomes of triple-negative breast cancer by applying iTRAQ technology. Background technique [0002] Triple-negative breast cancer (TNBC) refers to estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (human epidermal growth factor receptor 2). Receptor 2, Her2) are not expressed in a type of breast cancer, which has the characteristics of younger onset, high degree of malignancy, strong invasiveness, only part of which is sensitive to radiotherapy and chemotherapy, and has the characteristics of difficult treatment and easy recurrence. The peak period of recurrence is within a year, and most patients die in the first 5 years of treatment, which is one of the most difficult subtypes of breast cancer treatment. The pathogenic mechanism and interacting proteins of tri...

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Problems solved by technology

[0005] Then, can the application of iTRAQ technology to study the differentially expressed proteins in blood exosomes of patients with triple-negative breast cancer provide a reference for the disease mechanism of triple-negative breast cancer and the discovery of biomarkers related to triple-negative breast cancer? At present, there is no report on the method of using iTRAQ technology to study differentially expressed proteins in triple-negative breast cancer

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