Application of itraq technology to study the method of differentially expressed proteins in exosomes of triple-negative breast cancer

Abstract

The invention discloses a method for applying iTRAQ technology to study differentially expressed proteins of triple-negative breast cancer exosomes, comprising the following steps: (1) extracting and processing exosomes and exosome proteins; (2) performing step (1) The obtained exosomal protein sample is subjected to enzymatic hydrolysis and desalting to obtain a polypeptide; (3) iTRAQ labeling and mixing are performed on the polypeptide obtained in step (2); (4) HPLC separation and LC are performed on the mixture of polypeptide peptides labeled in step (3) ‑MS analysis; (5) Bioinformatics analysis and quantitative analysis were performed on the above-obtained mass spectrometry identification results to obtain differentially expressed proteins of triple-negative breast cancer exosomes. The iTRAQ technology provided by the invention combined with the MS / MS technology can quickly and effectively conduct the research on the exosome proteome of triple-negative breast cancer, and provide a reference for studying the disease mechanism of triple-negative breast cancer and discovering biomarkers related to triple-negative breast cancer.

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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