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Background: Various studies have verified the prognostic significance of the tumor-stromal ratio (TSR) in several types of carcinomas using manually assessed H&amp;E stained histologic sections. This study aimed to establish a computerized method to assess the TSR in invasive breast cancer (BC) using immunohistochemistry (IHC)-stained tissue microarrays (TMAs), and integrate the TSR into a novel nomogram for predicting survival. Methods: IHC-staining of cytokeratin (CK) was performed in 7 prepared TMAs containing 240 patients with 480 invasive BC specimens. The ratio of tumor areas and stromal areas was determined by the computerized method, and categorized as stroma-low and stroma-high groups using the X-tile software. The prognostic value of the TSR at 5-year disease free survival (5-DFS) in each subgroup was analyzed. Univariate and multivariate analyses were performed and a novel nomogram for predicting survival in invasive breast cancer was established and assessed. Results: The newly developed computerized method could accurately recognize CK-labeled tumor areas and non-labeled stromal areas, and automatically calculate the TSR. Stroma-low and stroma-high accounted for 38.8% (n = 93) and 61.2% (n = 147) of the cases, according to the cut-off value of 55.5% for stroma ratio. The Kaplan-Meier analysis showed that patients in the stroma-high group had a worse 5-DFS compared to patients in the stroma-low group ( P = 0.031). Multivariable analysis indicated that the T stage, N status, histological grade, ER status, HER-2 gene, and the TSR were potential risk factors of invasive BC patients, which were included into the nomogram ( P &lt; 0.10 for all). The nomogram was well calibrated to predict the probability of 5-DFS and the C-index was 0.817, which was higher than any single predictor. A dynamic nomogram was built for convenient use. The area under the curve (AUC) of the nomogram was 0.870, while that of the TNM staging system was 0.723. The Kaplan-Meier analysis showed that the nomogram had a better risk stratification for invasive BC patients than the TNM staging system. Conclusions: Based on IHC staining of CK on TMAs, this study successfully developed a computerized method for TSR assessment and established a novel nomogram for predicting survival in invasive BC patients.

Computerized Assessment of the Tumor-stromal Ratio and Proposal of a Novel Nomogram for Predicting Survival in Invasive Breast Cancer