Evaluation of alpha-methylacyl-CoA racemase, metallothionein and prostate specific antigen as prostate cancer prognostic markers.

Current diagnostic techniques are inefficient in distinguishing latent and low-risk forms of prostate cancer from high-risk forms. The present study is focused on determination of putative tumor markers of aggressive high-grade forms of prostate cancer. Potential markers were determined in blood sera of 133 patients (82 cases and 51 controls) and in cell lines (Gleason score 9-derived 22Rv1 and normal tissue derived PNT1A) on mRNA and protein levels. Alpha-methylacyl-CoA racemase (AMACR), metallothionein classes 1A and 2A (MT1A and MT2A) were determined and compared to prostate specific antigen (PSA) levels. On mRNA level, significantly increased expression of MT2A (2.4-fold), PSA (2.6-fold) and AMACR (8.4-fold) and insignificantly (1.9-fold) elevated MT1A in 22Rv1 compared to non-tumor PNT1A were determined. On protein level, significant enhancement of free PSA and total PSA in tumor cell line was evident. AMACR protein was 1.5-fold elevated in tumor line (below the level of significance). Contrary to mRNA, significantly (p = 0.01) reduced level of MT protein in tumor lines was determined. In the case of serum level, significantly enhanced MT level (4.5-fold) in patients' sera was found. No significant changes were observed in the case of AMACR. These findings indicate possible alternative role of MT to PSA prostate cancer marker. In addition, level of AMACR is distinctly higher in the Gleason score 9 in serum of patients and MT shows a descending trend in relation to Gleason score.

[Molecular mechanisms of zinc in prostate cancer]. / Zinek--molekulární mechanizmy u karcinomu prostaty.

In many developed countries, prostate cancer is the most common male tumour disease. The high incidence and mortality requires early diagnosis, differentiation of aggressive, highly malignant forms from clinically silent forms and understanding of the pathogenesis with its typical metabolic aberrancies (if any) in order to develop new targeted therapies. Prostate cells (including prostate cancer cells) are unique in their relation to zinc ions. Prostate tissue can accumulate these ions in up to tenfold higher concentration than other body cells. These ions influence many cellular processes incl. proliferation, differentiation and apoptosis. Prostate cancer cells lack ability to accumulate zinc. Therefore, zinc ions may be expected to play an important role in the disease pathogenesis, in its propagation and metastatic potential of tumour cells. Intracellular zinc levels are regulated by zinc-binding proteins, especially metallothioneins, and zinc transporters. Zinc level regulation dysfunction has been identified in prostate cancer cells and may thus play an important role in the prostate cancer pathogenesis. Moreover, due to its overproduction by prostate tissue, metallothionein serum levels are elevated and can be used as an important tumour marker.

[Hereditary pheochromocytoma--a family affected by von Hippel-Lindau disease]. / Hereditární feochromocytom--rodina postizená von Hippel-Lindauovou chorobou.

The authors present a case of a 37 year old male (proband) with a 13 year history of progressive sight impairment leading to blindness and a 4 year history of a mild hypertension. He was incidentally found to have large adrenal tumors after an ultrasound kidney examination. The tumors were confirmed with CT scan and magnetic resonance imaging. A bilateral pheochromocytoma was biochemically demonstrated and successfully removed. The eye diagnosis of angiomatosis retinae von Hippel-Lindau was ascertained after a search of the patient files in other medical departments, which led to a family screening. Proband's brother, having hypertension and a history of a cerebellar astrocytoma operation, was also diagnosed with CT scan to have a bilateral pheochromocytoma. Unfortunately, at the same time he was found to have a large irremovable neuroendocrine pancreatic carcinoma, which caused complications and his eventual death. Both proband and his brother were affected by the same CGG(Arg167)->CAG(Gln) mutation in the exon 3 of the VHL gene. Other living and examined family members were not affected, which was confirmed by negative genetic testing. One year after the pheochromocytoma operation, proband was diagnosed to have a retroperitoneal tumor left to the aorta, clinically silent, with slightly and non-constantly elevated urine norepinephrine and normetanephrine. Metaiodobenzylguanidine scintigraphy showed that it was a paraganglioma. The old CT and magnetic resonance picture review demonstrated that the tumor had already been present at the time of the operation. It was surgically removed and histologically verified. It is a pity that proband had not been sent by his ophthalmologist for an endocrine examination when the eye diagnosis was determined. Affection of the family would have been discovered earlier, and proband's brother might have possibly been saved.

Androgen receptor in prostate carcinoma: immunohistochemical and ligand saturation analyses.

Status of androgen receptor (AR) in prostate carcinoma is biologically important. Therefore, more methods assessing AR abnormalities are warranted. Immunohistochemical (IHC) and ligand saturation (LSA) assays were not compared in details. AR in 53 cases were tested by monoclonal antibody (Ab) F39.4.1 (Biogenex), polyclonal Ab N-20 (Santa Cruz) and by ligand saturation analysis with (3)H-methyltrienolon. Statistical analyses were performed with Spearman's nonparametric rank test including neoadjuvant therapy subgroups treated by antiandrogens, combined androgen blockade (22 cases; flutamide with gosereline) or without therapy. By using monoclonal Ab we found AR positive tumor nuclei in 46 cases. Mean of positives was 64%, median 75%. The polyclonal Ab was not sufficiently specific. With LSA AR were found in 43 cases. Mean level was 6.6 fmol/mg, median 5.5 fmol/mg. Comparing IHC with LSA, we noted correlation trend only for the monoclonal Ab (r=0.35; p=0.02). With thresholds 70% positive nuclei for IHC and 6 fmol/mg for LSA, there were 66% and 43% cases positive with IHC and LSA, respectively. The LSA and IHC positives did not show significant agreement, concordance level being 58% only. We found significant IHC-LSA correlation (r=0.68; p=0.004) solely in combined androgen blockade subgroup with 82% level concordance. Our study has demonstrated that AR IHC and LSA are independent complementary methods. Significant correlation between LSA and IHC show only cases treated with combined androgen blockade. An explanation hypothesis is discussed concerning LH-RH influence on free AR capable of ligand binding. IHC as well as LSA have specific biologic significance and may be useful for prostate cancer diagnostic and therapy.

Relation of prenephrectomy CD profiles and serum cytokines to the disease outcome and response to IFN-alpha/IL-2 therapy in renal cell carcinoma patients.

Immune parameters, including cytokine levels and CD profiles were determined in 78 renal cell carcinoma patients (RCC) prior to nephrectomy. The values were correlated with the outcome of disease and response to cytokine-based treatment during a 3-year follow-up. Significantly lower frequency of progressions and higher proportion of survivors were recorded in 24 treated patients compared to 43 untreated ones (22.9% vs. 53.5% and 82.9% vs. 55.8%) illustrating the beneficial effect of immunotherapy on the course of RCC at localized stage. RCC-related immune changes are demonstrated by reduced proportion of CD19+, CD28+, HLA-DR+, CD19+/80+ and CD8+/28+ subsets, by increased serum levels of IL-6, sIL-2R, CRP and by impaired production of IL-2 and TNF-alpha released by in vitro stimulated PBMC. Only increased CRP, IL-6 serum values, decreased CD8+ and increased CD122+ were significantly related to patients' prognosis. Comparisons of preoperative CD profiles and cytokine values with the response to IL-2/IFN-alpha based therapy disclosed significant correlation in only CD80+ and CD19+/80+ subsets. Treated patients who relapsed during the 3-year follow-up exhibited at the diagnosis significantly reduced proportion of CD80+ and CD19+/80+ cells (CD80+ means - 0.79 vs. 1.69 and CD19+/80+ means - 0.32 vs. 0.61) comparing to those surviving disease-free. In addition initial proportion of CD3+, CD8+ and CD19+ cells was reduced in treated patients who manifested progression but statistical difference from those remaining disease-free was not proved.

Renal cell carcinoma-associated immune impairment that may interfere with the response to cytokine therapy.

This prospective study was carried out to explore cytokine-related immune alterations in 69 renal cell carcinoma patients (RCC) and to look for changes which might potentially serve as a reliable predictors of response to cytokine-based therapy. Interleukin-2 (IL-2), its soluble receptor (sIL-2R) and tumor necrosis factor (TNF-alpha) levels produced in vitro by PHA activated and intact mononuclear cells (PBMC) were determined. Concentrations of IL-2, IL-4, IL-6, sIL-2R, TNF-alpha and CRP were measured in sera. Cytokine level was evaluated by enzyme-linked immunoadsorbent assay (ELISA) and CRP was determined by means of turbidimetric method. All measurements were performed in patients without any prior treatment. PHA activated PBMC of RCC patients were significantly defective in producing IL-2 and TNF-alpha comparing to controls (p < 0.03 and p < 0.001). The difference of sIL-2R was noted in metastatic stage only (p < 0.03). Unstimulated PBMC manifested decrease in IL-2 (p < 0.03) and increased level of TNF-alpha in advanced disease (p < 0.02). This impairment reflected tumor size and differentiation stage. Serum concentrations of IL-2, sIL-2R and TNF-alpha were within normal range. However, in relation to the clinical stage, significantly increased serum IL-2 was noted in combined Stage I and II as compared to controls (p = 0.012). IL-6 and CRP showed markedly elevated levels with a significancy which allowed to distinguish samples from metastatic patients. In conclusion careful comparisons of these data with clinical course of cytokine treated patients will disclose which of those tests may possess predictive power in the individual patients who are likely to respond to cytokine-based treatment.