POLYAMINES AS NEW POTENTIAL BIOMARKERS FOR DIFFERENTIAL DIAGNOSIS OF PROSTATE CANCER AND ESTIMATION OF ITS AGGRESSIVENESS.

BACKGROUND: Prostate cancer (PCa) is one of the most common malignancies in older men. The study of tissue markers of PCa can provide information about the state of proliferation and apoptosis in tumors, the susceptibility of tumor cells to metastasis and the mechanisms of resistance to therapy, which, in turn, can help predict the course of the disease and develop personalized treatment. Polyamines (PAs) spermine, spermidine, putrescine are of particular interest in terms of PCa tissue markers. AIM: To investigate the levels of basic and acetylated forms of PAs in the postoperative samples of malignant and benign tumors of the human prostate and evaluate the possibility of their use for differential diagnosis and assessment of the PCa aggressiveness. OBJECT AND METHODS: 57 postoperative tumor samples from patients with prostate adenocarcinoma of different Gleason score (GS) and clinical stage (T1-T4) and 20 samples of tumors from patients with benign prostate hyperplasia (BPH) were studied. The content of PAs was determined by high performance liquid chromatography. RESULTS: Among the studied PAs, the most significant difference between PCa and BPH was observed for spermine (Spm). The level of Spm in PCa samples was 16 times lower than in BPH samples (p < 0.01). We did not find a significant dependence of PAs levels, including Spm, on the clinical stage. The association between the Spm level and the GS was established. The indolent (GS6) tumors were characterized by the highest Spm level while in the most aggressive (GS9 and GS10) tumors Spm content was the lowest. CONCLUSIONS: A sharp decrease in Spm levels is probably a characteristic feature of prostate malignant tumors. The obtained results indicate an association of Spm levels in tumors with the GS. This may indicate Spm involvement in the formation of the aggressiveness of PCa. The results of the study can be further used for differential diagnosis of prostate tumors and for assessing the aggressiveness of PCa.

Application of gold and silver nanoparticles for selective assay of spermine in mixture with spermidine.

AIM: To assess the applicability of the novel technique based on the detection of spermine in solutions by spectrocolorimetric method using gold and silver colloidal nanoparticles. MATERIALS AND METHODS: Colloidal solution of gold nanoparticles were synthesized by chemical reduction of tetrachlorauric acid with trisodium citrate. Colloidal solution of silver nanoparticles was obtained by chemical reduction of silver nitrate with tryptophan. The absorption spectra of gold/silver metal colloids and their mixtures with polyamines were recorded. RESULTS: The increase of spermine concentration in solution caused the change in the intensity of the band of localized surface plasmon resonance that was not affected by the excess of spermidine. The color shift in colloidal gold due to its aggregation with spermine was registered spectrophotometrically. CONCLUSION: The principal possibility of selective quantification of spermine in the presence of spermidine in extremely high concentration using colloidal gold has been shown. This method can be used to assay selectively spermine in biological fluids.

Green tea, red wine and lemon extracts reduce experimental tumor growth and cancer drug toxicity.

AIM: To evaluate antitumor effect of plant polyphenol extracts from green tea, red wine lees and/or lemon peel alone and in combination with antitumor drugs on the growth of different transplanted tumors in experimental animals. MATERIALS AND METHODS: Green tea extract (GTE) was prepared from green tea infusion. GTE-based composites of red wine (GTRW), lemon peel (GTRWL) and/or NanoGTE as well as corresponding nanocomposites were prepared. The total polyphenolics of the different GTE-based extracts ranged from 18.0% to 21.3%. The effects of GTE-based extracts were studied in sarcoma 180, Ehrlich carcinoma, B16 melanoma, Ca755 mammary carcinoma, P388 leukemia, L1210 leukemia, and Guerin carcinoma (original, cisplatin-resistant and doxorubicin-resistant variants). The extracts were administered as 0.1% solution in drinking water (0.6-1.0 mg by total polyphenolics per mouse per day and 4.0-6.3 mg per rat per day). RESULTS: Tumor growth inhibition (TGI) in mice treated with NanoGTE, cisplatin or cisplatin + NanoGTE was 27%, 55% and 78%, respectively, in Sarcoma 180%, 21%, 45% and 59%, respectively, in Ehrlich carcinoma; and 8%, 13% and 38%, respectively in B16 melanoma. Composites of NanoGTE, red wine, and lemon peel (NanoGTRWL) enhanced the antitumor effects of cyclophosphamide in mice with Ca755 mammary carcinoma. The treatment with combination of NanoGTE and inhibitors of polyamines (PA) synthesis (DFMO + MGBG) resulted in significant TGI of P388 leukemia (up to 71%) and L1210 leukemia. In rats transplanted with Guerin carcinoma (parental strain), treatment with GTRW or GTE alone resulted in 25-28% TGI vs. 55-68% TGI in cisplatin-treated animals. The inhibition observed in the case of combination of GTE or GTRW with cisplatin was additive giving 81-88% TGI. Similar effects were observed when combinations of the cytostatics with GTE (or NanoGTE) were tested against cisplatin- or doxorubicin-resistant Guerin carcinoma. Moreover, the plant extracts lowered side toxicity of the drugs. Treatment with GTE, NanoGTE, and NanoGTRW decreased the levels of malondialdehyde in heart, kidney and liver tissue of experimental animals, as well as the levels of urea and creatinine in blood serum, increased erythrocyte and platelet counts, hemoglobin content, and decreased leucocyte counts. CONCLUSION: The obtained data indicate the prospects for further development of GTE and corresponding nanocomposites as auxiliary agents in cancer chemotherapy.

Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis.

AIM: To study the influence of polyamine metabolism inhibitors on the growth, metastasis and ornithine decarboxylase (ODC) activity of Lewis lung carcinoma. MATERIALS AND METHODS: Experiments were performed on female mice C57Bl/6 with Lewis lung carcinoma. Nω-hydroxy-nor-arginine (nor-NOHA) and α-difluoromethylornithine (DFMO) were used as arginase and ODC inhibitors, correspondently. Inhibition of tumor growth was calculated by comparison of tumor volume in the treated and control groups. The average number of metastases per animal in the group and the average volume of pulmonary metastases per animal in the group have been determined. Determination of ODC - the key enzyme of the polyamine synthesis - in the samples of experimental tumors was performed by method of Luqman S. RESULTS: Administration of DFMO or it's combination with nor-NOHA resulted in the decrease of tumor growth rate, number and volume of lung metastases and was accompanied with reduced ODC activity in tumor tissue. CONCLUSION: Modifiers of polyamine metabolism may be considered as promising targeted cancer therapy.

Relationship between NF-κB, ER, PR, Her2/neu, Ki67, p53 expression in human breast cancer.

AIM: The aim of the present study was to investigate expression patterns of transcription factor NF-κB (p50 and p65), ER, PR, Her2/neu, Ki-67 and p53 in tumor tissue of patients with breast cancer (BC) and analyze correlation between these markers. PATIENTS AND METHODS: 62 BC patients previously not treated with chemo- or radiotherapy were included in the study. All tumors belong to invasive ductal carcinoma of different grade. Expression of molecular markers was determined by immunohistochemical analysis on paraffin-embedded tissue sections. RESULTS: The correlation between tumor grade and expression of ER, PR, Ki-67 and p53 was defined. NF-κB expression was found to be changed dependent on expression of ER, PR and p53 and also on molecular subtype (luminal, Her2-positive, hybrid, basal-like). The highest levels of NF-κB, Ki-67 and p53 were found in Her2/neu+ and basal-like tumor subtypes. CONCLUSION: The increase of nuclear expression of NF-κB correlates with a decrease of expression of steroid hormone receptors (ER and PR), increase of p53 accumulation, and is associated with Her2-positive and basal-like tumor types.

[The biological and diagnostic implication of the comparative data on expression of the same protein, obtained by Western blotting and surface plasmone resonance tests].

The comparative analysis of the data on the expression of the same modulating protein MX, obtained on the same biological materials by Western blotting and surface plasmone resonance tests was shown to provide in most cases a semiquantitative opinion as to modulation of MX affinity to its specific target (effector X protein) under the experimental action of F. This is doubtlessly to offer additional possibilities for the prediction and differential diagnosis of a number of diseases, including cancer.

Effects of green and black tea biocomposites on endogenous synthesis, metabolism and genotoxic effect of carcinogenic N-nitrosodimethylamine.

AIM: To study the modifying effect of green and black tea biocomposites on endogenous synthesis and genotoxic action of the carcinogenic N-nitrosodimethylamine. METHODS: Green and black tea biocomposites were administered to the white inbred rats in vivo. Amidopyrine and sodium nitrite were used as N-nitrosodimethylamine precursors and 4-methylpyrazol as an inhibitor of its metabolism. N-nitrosodimethylamine (blood, daily urine and reaction mixture), nitrites and nitrates (daily urine) levels were measured. Genotoxic action was tested by formation of DNA single-strand breaks in hepatocytes. RESULTS: In in vitro system, biocomposites increased N-nitrosodimethylamine synthesis in neutral medium and decreased in acid conditions. In vivo, black tea biocomposite consumption resulted in enhanced background level of DNA single-strand breaks in rats hepatocytes and higher genotoxic effect upon administration of N-nitrosodimethylamine precursors. The levels of N-nitrosodimethylamine in blood and urine of experimental animals were increased after precursors' administration. In contrast, green tea biocomposite significantly decreased background level of DNA single-strand breaks. However, there was no protective action of this food supplement at the N-nitrosodimethylamine, precursors' administration. 4-methylpyrazol administration did not increase N-nitrosodimethylamine excretion in urine, while this effect was observed in control and black tea biocomposite groups. CONCLUSIONS: The effects of green tea and black tea biocomposites on N-nitrosodimethylamine synthesis in in vitro system are unidirectional and depend on biocomposites' concentration and acidity of the medium. Long-term consumption of black tea biocomposite resulted in intensification of endogenous N-nitrosodimethylamine synthesis and increased damage of the hepatocytes' DNA. As to the green tea biocomposite, the obtained results allow us to suggest that this biocomposite enhanced N-nitrosodimethylamine metabolism.

[Polyamines and mental diseases].

The authors make an attempt to understand and evaluate the role of polyamines in the development of endogenous mental diseases. The article is dedicated to distribution and exchange of polyamines in the central nervous system, their metabolic and regulatory associations with gamma-aminobutyric acid and dopaminergic systems, as well as to possible neuromediatory and neuromodulatory functions of polyamines. The paper is also concerned with effects of psychotropic agents on polyamine metabolism and contains a hypothesis on the role of polyamines in etiopathogenesis of schizophrenia.

[Effect of alpha-difluoromethylornithine and polyhexamethyleneguanidine (PMG), polyamine biosynthesis inhibitors on leucosis L1210 growth kinetics and life expectancy of animals with cancer].

The article deals with the study of polyamines content and y-glutamiltranspeptidase (gamma-GTP) activity in leucosis L1210 cells under the influence of inhibitors of polyamines synthesis such as alpha-difluoromethylornithine (alpha-DFMO) and polyhexamethylenguanidine (PMG). Injections of alpha-DFMO and PMG to animals essentially reduce putrescine and spermidine concentrations, and the levels of spermine and gamma-GTP activity increase under this influence. These modulation were associated with L1210 leucosis growth retardation. Antiblastic effect was dependent on inhibitors' doses and mode of injections' course. Under the optimum conditions the retardation index was 90-98%. The animals with retarded tumor growth had essentially longer survival time frame than blank tumor-bearing animals (index was 37.2 for a-DFMO and 67.5 for PMG).

Ornithine decarboxylase activity and polyamine content in adenocarcinomas of human stomach and large intestine.

Ornithine decarboxylase (ODC) activity and differential polyamine composition have been studied in macroscopically normal mucosa, adjacent mucosa, and neoplastic tissue of 95 patients with adenocarcinomas of stomach and large intestine. In tumors, we found increased ODC activity and polyamine content as compared with surrounding mucosa. ODC activity in macroscopically normal tissue of patients with tumors of stomach and large intestine increased as the disease progressed. An inverse relationship was observed between ODC activity in adenocarcinomas and differentiation.

[Antimetastatic effect of L-lysine-alpha oxidase]. / Antimetastaticheskii éffekt L-lizin-alfa-oksidazy.

The influence of a new antitumor enzyme L-lysine alpha-oxidase on Lewis lung carcinoma spreading was studied in mice in which primary tumor had been removed. The enzyme was found to significantly decrease the extent and number of lung metastases as compared to mice which hadn't received L-lysine alpha-oxidase. This was matched by recovery of alveolar macrophages functional activity, as assessed by adenosine deaminase and 5' nucleotidase levels in these cells. Moreover, antimetastatic and cytostatic effect was confirmed by the measuring of polyamine concentration in mice erythrocytes.

[Ornithine decarboxylase activity and polyamine contents in 1,2-dimethylhydrazine-induced carcinogenesis of the intestines in rats]. / Aktivnost' ornitindekarboksilazy i soderzhanie poliaminov pri indutsirovannom 1,2-dimetilgidrazinom kantserogeneze kishechnika u krys.

The ornithine decarboxylase (ODC) activity and concentration of polyamines have been studied in small and large intestine during 1,2-dimethylhydrazine-induced carcinogenesis in rats. Changes in the polyamine biosynthesis consisting both in enhanced ODC activity and an increase of the intracellular content of putrescine, spermidine and spermine. Process of the polyamine synthesis activation proceeds in two phases: the most expressed and similar changes in polyamine metabolism factors have been observed at early (the 1st month) and late (the 5th-6th months) stages of carcinogenesis. It is supposed that intensification of the polyamine synthesis is a typical feature of malignization in the gastrointestinal tract.

[Effect of cyclophosphane on polyamine excretion in rats with Guérin's carcinoma and Pliss' lymphosarcoma]. / Vliianie tsiklofosfana na ékskretsiiu poliaminov u krys s kartsinomoi Gerena i limfosarkomoi Plissa.

Polyamines were determined in the urine of rats with the Guerin carcinoma and Pliss lymphosarcoma treated with cyclophosphamide. Alterations in polyamine distribution and rise of the spermidine/putrescine ratio were caused by the tumor regression or partial inhibition of the tumor growth. Regression of the Guerin carcinoma was accompanied by a rise of spermidine concentration in urine. Delay of the Pliss lymphosarcoma growth led to the putrescine level stabilization. Exposition of the rat normal tissues and cells to the cytostatic caused the alteration in the urinary polyamine excretion. These data suggests that the polyamine test may be useful to evaluate the efficiency of the anticancer treatment soon after its beginning.

[Polyamines of lymphocytes in animals in an experimental tumor process]. / Poliaminy v limfotsitakh zhivotnykh pri éksperimental'nom opukholevom protsesse.

Total content and individual fractions of polyamines were studied in lymphocytes of thymus and spleen tissues as well as in lymphatic glands of mice with Lewis carcinoma (strain C57BL/6). Fraction composition as well as total content of polyamines were altered during malignization, as a result of which proliferative activity of lymphocytes was shifted.

[Increase in ornithine decarboxylase activity and polyamine levels of the rat liver during an early period of hepatocarcinogenesis]. / Uvelichenie aktivnosti ornitindekarboksilazy i soderzhaniia poliaminov v pecheni krys v rannii period gepatokantserogeneza.

The ornithine decarboxylase activity and the polyamine content in the fraction of plasma membranes and cytosol of the rat liver are studied in the early period (1-4 weeks) of nitrosodiethyl amine-induced hepatocarcinogenesis. The enzyme activity and polyamine levels in the cytosol of the rat liver cells are found to increase sharply during the first month of the disease, the maximum being observed the first-second week. By the end of the fourth week these indices become lower but they remain significantly higher than the normal levels. The polyamine level increases considerably in the fraction of plasma membranes with the maximum observed the second week.

[Ornithine decarboxylase activity and polyamine content in human adenocarcinomas of the stomach and large intestine]. / Aktivnost' ornitindekarboksilazy i soderzhanie poliaminov v adenokartsinomakh zheludka i tolstogo kishechnika cheloveka.

Ornithine decarboxylase (ODC) activity and polyamine content were studied in the intact mucosa and tumours of human stomach and large intestine adenocarcinomas. ODC activity has been found to be much higher in the neoplastic tissue than in the corresponding intact mucosa. Concentrations of polyamines in neoplasms were also higher than in mucosa. Interrelation was found between ODC activity polyamine content and degree of differentiation of the tumour cells in rectal adenocarcinomas.

[Diagnostic and prognostic importance of the polyamine test of patients with ovarian tumors]. / Diagnosticheskoe i prognosticheskoe znachenie poliaminnogo testa u bol'nykh s opukholiami iaichnikov.

Assays of urine-polyamine levels were run in 66 patients with ovarian tumors. The levels of polyamine excretion were found to be significantly higher in cases of malignant epithelial tumors than in control. In the course of effective therapy, polyamine concentration showed a sharp increase followed by a decrease in posttreatment period. The index of therapy efficacy (Formula: see text) was in a strict correlation with the clinical status in 78% of cases. Polyamine tests should be recommended for diagnosis, evaluating the effectiveness of therapy and making prognosis.

[Various aspects of the biological role of polyamines in normal and malignant growth]. / Nekotorye aspekty biologicheskoi roli poliaminov pri normal'nom i opukholevom roste.

The data on location of polyamines (spermine, spermidine, putrescine) in cells of different tissues and organs, intracellular polyamines distribution and the factors controlling polyamine metabolism are reviewed. Certain aspects of a regulatory role of polyamines in nucleic acid and protein synthesis and of changes in the activity of the enzymes participating in the synthesis and degradation of polyamines in the neoplastic growth are dealth with. The data available in literature and the results of the authors own investigations concerning the use of the polyamine test for diagnosis and evaluation of malignant tumours treatment efficiency are presented.