Proteomic Analysis Of Breast Tumor By DIA Discovers Two Metabolic Proteins With Tumor Aggression

Breast cancer is the second most frequent cancer after lung cancer,causing a great threat to women's life.In the meantime it causes a great financial burden to the society.Therefore,the study of the pathogenesis of breast cancer has a great significance.Tumor research has been going on for many years,and breast cancer-related-omics research has extensively explored genomes,transcriptomes,metabolism and so on.We use proteomics as the starting point because protein has the most direct contact with life activity.There are a lot of methods to study proteomics,such as DIA,PRM,TMT,ITRAQ.DIA is a label free protein quantitative method,this method can be used with a special sample for multi-channel.This method is more economical compared with TMT and ITRAQ labeling methods.This experiment uses a MC10AT cell model,which simulates the pathogenesis of breast cancer.We first used the DIA method to carry out the proteomics study of the cell model,identified more than 3000 proteins,and identified 19 proteins using PRM for validation.Through our PRM data analysis,it was found that 13 proteins were indeed significantly different in CAla and CAlh.At the same time,we also used the QPCR method to verify the target protein from the mRNA level.In order to further study the function of the target proteins we selected two proteins(ACLY--ATP Citrate Lyase and FASN--Fatty Acid Synthase)and constructed plasmids to knock down the target gene in the MCF10CAla cell line.After confirming the knockdown efficiency,the effect of the target genes on the migration of tumor cells was confirmed by in vitro cell migration experiments.