Ben Jemaa et al. Journal of Experimental & Clinical Cancer Research 2010, 29:171 http://www.jeccr.com/content/29/1/171 Page 4 of 9 Figure 1 H & E stained slides in NP (B), BPH (F) and PC (J); immunohistochemical localizations of PSMA, PSA and CD34. Negative control (A). NP showing weak cytoplasmic staining for PSMA (C) and PSA (D) in epithelial cells. CD34 was found at low level in membranous and cytoplasmic endothelial cells in NP (E) and BPH (I). BPH showing weak membranous staining for PSMA (G) and strong membranous and cytoplasmic staining for PSA (H) in prostatic epithelial cells. PSMA (K) and CD34 (M) showed strong immunoreactions in infiltrating prostatic carcinoma. PSA (L) showed weak cytoplasmic immunoreactions of epithelial cells in PC. Scale bars: A-G, I-M, 20 μm; H, 30 μm. samples, we divided BPH and PC samples into 3 sub- groups. The first group has a CD34 OD values between 2.34 and 8, the second group has a CD34 OD values between 8 and 12.08 and the third group has a CD34 OD value superior to 12.08 (Figure 2C and Figure 3). In BPH samples, no difference neither in PSA nor PSMA expression was found in all 3 subgroups (Figure 3A). Importantly, depending on the degree of vascularisa- tion, we found an inverse relation between angiogenesis and PSA in PC patients. Unlike PSA, the highest intra- tumoral angiogenesis is accompanied by high PSMA expression in prostate cancer cells (Figure 3B). To study the distinct pattern of proteins tumour profiles produced by prostate epithelial cells we established differ- ent prostate-associated antigens profiles depending on positive immunoreactions to PSA and PSMA in NP, BPH and PC samples. We obtained a negative group for PSA and/or PSMA in each prostate type. The distribution of this group was as followed: 2 in NP, 13 in BPH and 11 in PC patients. Figure 4 showed 4 prostate-associated antigen profiles expressed differently in NP, BPH and PC patients as followed: (PSA+, PSMA+), (PSA+, PSMA-), (PSA-, PSMA-) and (PSA-, PSMA+). For all histological speci- mens, the profile (PSA+, PSMA+) was the most expressed in 66% of NP, 70% of patients with BPH and 71% of PC patients. However, no significance was observed between the different groups of prostatic specimens according to the percentage of immunoexpression of the profile (PSA+, PSMA+). To obtain insights into the relationship between PSA and PSMA production in the subgroup (PSA+, PSMA+) along prostatic diseases, we analysed the intensi- ties of immunoreactions to PSA and to PSMA in NP, BPH and PC patients for the above profile. As observed in Figure 5, optical density of PSA increases significantly from NP to BPH and declines in PC samples in the profile (PSA+, PSMA+) (p < 0.0001). However, the intensity of immunoreaction to PSMA increases significantly from NP to BPH and malignant prostate specimens (p < 0.0001) in the same profile. The prostate tumour profile (PSA+, PSMA-) expres- sion levels decreases from NP to benign prostatic tissue and primary prostate cancer (50% vs. 15% vs. 2%, respectively). Inversely, the profile (PSA-, PSMA+) expression increases from NP to BPH and PC patients