Hbre duc140sur 01

Breast cancer tissue section containing HBre-Duc140Sur-03 (140 cases) and HBre-Duc140Sur-01 (140 cases) were provided by Outdo Biotech (Shanghai, China). Histological parameters were performed in accordance with the criteria of the World Health Organization. Pathologic staging was determined by the current International Union against Cancer tumor-lymph node metastasis classication. The follow-up information of breast cancer patients were as follows: the operation time of these patients was from January 2001 to August 2004 and the final follow-up was July 2013, thus the follow-up range was 9–12.5 years. Among this follow-up time, 73 out of 278 patients died of breast cancer with a median OS time of 62 months (2–147 months), and 205 patients were still alive with a median OS time of 123 months (40–150 months). The clinical information of breast cancer patients were shown in Table 1. A total of 278 cases are included in the final analysis except for 2 cases missed clinical staging information.

Tissue sections from the in vivo experiments were used to detect Ki-67 and PCNA protein expression levels using immunohistochemistry. The indirect streptavidin-peroxidase method was used according to the manufacturer’s introduction. Immunohistochemically stained tissue sections were examined separately by two pathologists. The antibodies used were rabbit anti-PCNA (Cat. No.10205-2-AP, 1:30, Proteintech), anti-Ki67 (Cat. No. Ab16667, 1:100, Abcam).
Paraffin tissue arrays of breast cancer (HBre-Duc140Sur-01) was purchased from Shanghai Outdo Biotech (Shanghai Outdo Biotech Co., Shanghai, China). Tissue sections stained immunohistochemically for MAP2K4 and Vimentin were reviewed, and cytoplasmic staining scored separately by two pathologists blinded to the clinical parameters. The extent of staining, defined as the percentage of positively staining tumour cells in relation to the whole tissue area, was scored on a scale of 0-4 as follows: 0, <10%; 1, 10-25%; 2, 26-50%; 3, 50-75%; and 4, >75%. The staining intensity was scored as 0-3(Negative:0; Weak expression: 1; Positive expression:2; Strong expression:3; The sum of the staining intensity and staining extent scores was used as the final staining score for MAP2K4 and Vimentin(0-7). For statistical analysis, final staining scores of 0–5 and 6–7 were considered to respectively show low and high expression.