Fig. 1
Steps of experimental design and aspects of in vivo development of 4T1 breast carcinoma in BALB/c mice. (A) Experimental design of cancer induction with 1 × 106 4T1 cells representing the experimental design to step 1 to monitor evolution of 4T1 breast carcinoma and metastases and step 2 corresponding to tissue collection to qPCR experiments after determining the best collect point. (B) Clinical aspects of the evolution of 4T1 breast carcinoma in BALB/c mice including survival rate, clinical score over 30 days after injection of cells, assessment of tumor growth (mm3) over 30 days after injection of cells and evolution of ulcerated areas in the primary tumor over 30 days after injection of cells. (C) Scintigraphic images and biodistribution of 99mTc-HYNIC-βAla-Bombesin(7−14) in different tissues. The red arrow indicates renal clearance of peptide radiolabeled. The yellow arrow indicates an exclusion area of intense bladder uptake. The red circle indicates the primary tumor area. The yellow circle indicates an area without a tumor. The orange circle indicates the liver uptake area. (D) Percentage of injected dose per tissue gram in ex vivo experiments in liver, lung, and brain at 14, 21, and 28 days after injection of cells. In B, the log-rank test (Mantel-Cox) was used in the survival curve, and to calculate the Hazard Ratio, the Mantel-Haenszel test was used between the control group (n = 6) and animals with 4T1 breast carcinoma (n = 15); Mann-Whitney test was used in the clinical score, tumor growth and evaluation of ulcerated areas. The one-way ANOVA test, followed by the Tukey test, was used to analyze statistical differences between the control group (n = 6) and 4T1 group (n = 6) in D. Data are represented as mean ± standard error of the mean (SEM). Significant differences are represented by *p < 0.05; **p < 0.01; ***p < 0.001.
