Visualization And Prognostic Efficacy Of Breast Tumor Necrosis Based On Multiphoton Microscopy

Breast cancer is one of the most common malignant tumors in women,which seriously affects women’s physical and mental health and even endangers their lives.Breast cancer overtook lung cancer as the most common cancer in 2020.Tumor necrosis(TN)is a common histological feature of solid tumors,and its prognostic significance has been studied in several solid tumors including renal cell carcinoma aggressiveness,pancreatic ductal carcinoma,malignant mesothelioma,soft tissue sarcoma,hepatocellular carcinoma,cell carcinoma of the upper urinary tract,lung cancer and colorectal cancer.In these studies,there were two classification methods of TN,the presence/absence-based classification and extent-based classification.Based on the two classification methods,the studies indicated that the TN was associated with poor outcome.Similarly,the relationship between TN and IBC outcome has also been widely studied though these two necrotic classification methods.However,there were several conflicting results about the impact of TN on IBC prognosis.So,whether TN is an independent prognostic factor and its effect on staging remains unclear.Multiphoton microscopy(MPM)based on nonlinear signal inherent source is very suitable for high resolution imaging of tumor microenvironment.Since tumor cells contain NAD(P)H,flavins,retinol,tryptophan and its indoleamine derivatives,which would generate two-photon stimulated fluorescence(TPEF)signals,the location of cells could be clearly displayed by TPEF imaging without labelling.Collagen fibers have a non-centrosymmetric molecular structure and can be excited to generate second-harmonic generation(SHG)signals,that is,collagen morphology can be directly visualized in situ by SHG imaging.Due to the good ability to identify collagen morphology and location,SHG signal has advantages in analyzing stroma in tumor microenvironment.The relative spatial distributions of cells and collagen fibers in the tumor microenvironment could be well displayed in the MPM image.Therefore,in this study,MPM images of IBC patients’ microenvironment were obtained based on multi-photon microscopic imaging,and TN was divided into TN1-4according to the relative spatial positions of TN,tumor cells,collagen fibers and myoepithelial cells.According to the frequency of TN1-4 in the microenvironment,TN-Score of each patient was obtained,and a risk prognosis model was established according to TN-score.This is a new TN classification method to study the prognosis of IBC,which is different from traditional TN classification based on presence/absence and degree.Further,we included TN score into the current staging category to study the effect of TN on IBC staging.