THE CHANGES OF NO LEVEL AND RNase ACTIVITY IN TUMOR TISSUE ACCOMPANYING THE PROGRESSION OF PROSTATE CANCER.

AIM: To assess the inducible NO-synthase activity and the total RNase activity in tissue samples and blood neutrophils of the patients with prostate intraepithelial neoplasia (PIN) and prostate cancer (PCa) of different stages. MATERIALS AND METHODS: NO level was measured in tumor tissue and neutrophils of patients with PIN and PCa of different stages by electron paramagnetic resonance using the spin traps technology. RNase activity in tumor tissue of patients with PIN and PCa was measured by the method of zymography. RESULTS: We have found that NO levels in prostate tumor tissue were significantly higher than in the PIN and increased along with the disease progression. Analysis of NO level in neutrophils of the PCa patients demonstrated that the values were not dispersed and did not depend on the stage of disease. NO level in neutrophils of the PCa patients increased manifold as compared with that in healthy donors. At the same time, the RNase activity in the prostate tumor tissue gradually decreased with PCa progression. CONCLUSION: Activities of inducible NO-synthase and RNases change significantly with progression of PCa.

Expression of micro-RNA hsa-miR-30c-5p and hsa-miR-138-1 in renal cell carcinoma.

AIM: To analyze the expression levels of hsa-miR-30c-5p and hsa-miR-138-1 in tumors of patients with renal cell carcinoma to determine whether they could be used as diagnostic markers. MATERIALS AND METHODS: The relative expression of hsa-miR-30c-5p and hsa-miR-138-1 was compared in the paired samples of kidney tumor tissue and conventionally normal tissue adjacent to the tumor. RESULTS: We found a significant decrease in miR-30c-5p and miR-138-1 levels in tumor tissues even in the cases of early stage cancer. In addition, miR-138-1 expression was lower in renal cell carcinoma Fuhrman grade G3 + G4 as compared to Fuhrman grade G2. However, we found no association between miR-30c-5p and miR-138-1 expression in the tumors and the major clinical and pathological characteristics of renal cell carcinoma patients. CONCLUSIONS: A significant reduction in the expression levels of hsa-miR-30c-5p and hsa-miR-138-1 in renal cell carcinoma indicates the feasibility of further studies on the probable diagnostic utility of these markers.

[BIOMARKERS OF EARLY AND DIFFERENTIAL DIAGNOSIS OF PROSTATIC CANCER].

There were examined 87 patients, suffering prostatic gland diseases, in whom the level of general prostate-specific antigen (PSA) in the blood have constituted 0.5 ­ 30 ng/ml. In 27 of them prostatic gland diseases were not revealed, in 28 ­ prostatic cancer was diagnosed, and in 32 ­ benign prostatic hyperplasia. There was proved, that rate of prostatic cancer and benign prostatic hyperplasia have correlated with prostatic gland volume and depended upon PSA and its isoforms concentration in the blood. The [-2] proPSA in the blood serum level have differed in all three groups, and the level of general and free PSA ­ in patients of group I, comparing with that in the groups 2 and 3. That have witnessed the possibility of determining of the [-2]proPSA content as alternative biomarker of early and differential diagnosis of prostatic cancer.

[IDENTIFICATION OF A NEW DIAGNOSTIC MARKERS OF PROSTATIC CANCER, USING NOTI-MICROCHIPS].

The biopsy material specimens were investigated in 33 patients, examined for the prostatic cancer suspicion. In accordance to the morphological investigation data, in 15 patients a benign prostatic hyperplasia was verified, and in 18--pancreatic adenocarcinoma. NotI-Microchips of 180 clones of the third chromosome were used for determination of epigenetic changes. In 50 genes of the third chromosome a high rate of the methylation state changes (from 33 to 82%) was noted. Some changed genes take part in cancerogenesis (HMGB1L5, LRRC58, GPR149, DZIP1L, C3orf77, NUDT16) and in the prostatic gland cancer occurrence (BCL6, ITGA9, FBLN2, SOX2, LRRC3B etc.). Dependence of the genes methylation state from the clinic-morphological indices in patients with the prostatic gland cancer, including, the prostate-specific antigen level, the tumor differentiation degree in accordance to Gleason, was not established. Panel, consisting of 16 new potential markers for early and differentiated diagnosis of prostatic gland cancer, was identified: BHLHE40, FOXP1, LOC285205, ITGA9, CTDSPL, FGF12, LOC440944/SETD5, VHL, CLCN2, OSBPL10/ZNF860, LMCD1, FAM19A4, CAND2, MAP4, KY and LRRC58.

[PROGNOSTIC STRATIFICATION OF CLINICALLY LOCALLY SPREAD CANCER OF PROSTATIC GLAND].

In 2002 ­ 2015 yrs in the clinic radical prostatectomy was performed in 403 patients for prostatic cancer, including those having stage cТ3 ­ in 102 (25.3%). Basing on analysis of main risk factors, a model of prognostic stratification of clinically locally spread prostatic cancer on three groups was created: with low risk ­ in the absence of data, confirming invasion into seminal vesicles (stage cТ3а), the prostate­specific antigen (PSA) content less than 20 ng/ml, degree of cellular differentiation of tumor (Glisson's index) in accordance to biopsy data ­ 6 and less; with intermediate risk ­ while presence of one of unfavorable prognostic factors (stage cТ3b), PSA 20 ng/ml and more, Glisson's index 7 and more; with high risk ­ while presence of two and more unfavorable prognostic factors. The follow­up duration was 50,6 mo at average. Biochemical recurrence in the groups while low, intermediate and high risk presence have occurred, accordingly, in 14.3, 37.1 and 62.3% patients (р<0.05), and its occurrence risk in patients while intermediate risk present is in 3,5 times, in high risk ­ in 9.9 times bigger, than in a low risk. Stratification may appear useful for practical physicians and investigators while choosing tactics of treatment in patients with prostatic cancer clinical stage ІІІ

Correlation between histological type and immunohistochemical profile of prostate cancer and gleason scale gradation.

AIM: To evaluate the characteristics of prostate cancer (PC) morphogenesis, taking into consideration the role of proliferation and apoptosis in tumor cells. METHODS: p53, p16INK4a, Bcl-2 and Ki-67 proteins expression was analyzed by immunohistochemistry in paraffin embedded sections of biopsy specimens from PC patients. The level of tissue immunoreactivity was evaluated by semi-quantitative method with estimation of 100% colored cells content over 1000 cells in one specimen. Patients were divided into three groups in accordance to Gleason scale gradation: group 1 - with Gleason scale < 5 (n = 13); group 2 - with Gleason scale > 5 and < 8 (n = 8); group 3 - with the highest Gleason scale > 8 (n = 6). RESULTS: Upon histological examination of prostate biopsy specimens, it was found that in the first group in 6 out of 13 (46%) cases small acinic cell PC developed on the background of chronic prostatitis with PIA (proliferative inflammatory atrophy) locus, frequently in combination with prostatic intraepithelial neoplasia (PI) locus. Hyperchromic epithelial cells in PIA locus were characterized by nuclear expression of p53 and Ki67 proteins, and cytoplasmic expression of Bcl-2. The precancerous foci in the PIN and PIA in the biopsy specimens of the second group of PC patients were found in 2 out of 8 (25%) cases of large and small acinic cell adenocarcinoma observations. The expression level of p53, p16INK4a, Bcl-2 proteins and especially Ki67 protein adequately increased in tumors of group 2 in comparison with group 1. Group 3 comprised of patients with Gleason scale > 8, predominantly solid structures or scirrhus of PC, which were characterized by the highest nuclear expression of p53, p16INK4a and Ki-67, and also by overexpression of cytoplasmic Bcl-2. CONCLUSIONS: Obtained results showed the direct correlation between patients' Gleason scale, and the expression level of p53, p16INK4a, Bcl-2 proteins and, particularly, Ki67 marker of proliferating cells in PC tumor cells.