HER-2-positive patients were defined according to the criteria of Osman et al. [6] as 2+ and 3+ staining in more than 10% of PSA-positive cells. A mean expression of HER-2 per cell was calculated using the following formula: sum of HER-2 scores/number of cells Rapamycin IC50 counted.Samples were analyzed at low power, and photographed at a magnification of 400x using a digital camera, Samsung Digimax D73, and processed with the Digimax program for Windows 98. The immunocytochemical evaluation was performed by a single person, blinded to the clinical details using a coded system.The patients were divided into 3 groups: preradical prostatectomy and bone scan negative: patients with micrometastasis were analyzed for MMP-2 and HER-2 expressions, postradical prostatectomy bone scan negative without evidence of biochemical failure, defined as a PSA > 0.
2ng/mL and without androgen blockade: patients were analyzed for MMP-2 and HER-2 expressions, and postradical prostatectomy, biochemical failure, and with androgen blockade. Patients were analyzed for MMP-2 and HER-2 expressions, according to serum PSA at the time of sampling and time elapsed from starting of androgen blockade.2.4. Statistical AnalysisDescriptive statistics were used for demographic variables, expressed as mean and standard deviation in the case of continuous variables with a normal distribution. In case of an asymmetrical distribution the median and interquartile range (IQR) values were used. Noncontiguous variables were presented as frequencies.
The Student’s t-test was used to compare continuous variables with a normal distribution, Chi-squared, Kruskal-Wallis, and log regression for the differences in frequency. The kappa test was used for tests of concordance.The analysis was firstly to compare the expressions of HER-2 and MMP-2 in the micrometastasis and stromal expression of MMP-2 in men with and without androgen blockade and secondly, in the men undergoing androgen blockade, to compare the expressions of HER-2 and MMP-2 in the micrometastasis and stromal expression of MMP-2 with the serum PSA at the time of sampling and with the length of androgen blockade at the time of sampling.2.5. Ethical ConsiderationsThe study was directed with complete conformity with the principles of the declaration of Helsinki and approval of the local ethical committees.3. ResultsA total of 191 men participated in the study.
3.1. Preprostatectomy Radical35 men with a mean age of 70.0 �� 10.5 years and a median serum PSA of 4.83ng/mL (IQR 3.03�C11.48ng/mL) comprised the group. Overall micrometastasis was detected in 26/35 (74.3%); there were significantly fewer micrometastases detected in patients with Gleason 4 and stage 1 cancer (Table 1).Table 1Detection of micrometastasis (mM) according to Entinostat Gleason score and stage.MMP-2 expression was seen in 3/26 (11.