http://dspace.nbuv.gov.ua:80/handle/123456789/133124 2026-04-18T23:16:11Z http://dspace.nbuv.gov.ua:80/handle/123456789/138607 Genetic and epigenetic changes of NKIRAS1 gene in human renal cell carcinomas Gerashchenko, G.V.; Bogatyrova, O.O.; Rudenko, E.E.; Kondratov, A.G.; Gordiyuk, V.V.; Zgonnyk, Y.M.; Vozianov, O.F.; Pavlova, T.V.; Zabarovsky, E.R.; Rynditch, A.V.; Kashuba, V.I. Renal cell carcinoma (RCC) is the most common malignant tumor of kidney associated with the worst clinical outcome. No molecular markers for RCC diagnostics and prognosis that could be applied in clinics were described yet. Large-scale screening of 3p human chromosome genes/loci in RCC and histologically normal tissues surrounding the tumors using NotI-microarray approach demonstrated that NKIRAS1 gene contained the largest percent of genetic/epigenetic changes in RCC tumor cells. Aim: To validate the results of NotI microarray analysis and study genetic, epigenetic changes, and the expression level of NKIRAS1 gene in human RCC samples. Methods: DNA and RNA were isolated from freshly-frozen renal tumors’ samples (n = 12) and from normal tissues surrounding the tumors. Epigenetic changes (methylation status) of NKIRAS1 were detected by bisulfite sequencing. Genetic changes and expression level were analyzed by Quantitative real-time PCR (qPCR) with SYBR Green. For relative quantification 2-ΔΔCP method was used. Nonparametric tests (Wilcoxon, Kruskal — Wallis and Mann — Whitney) were applied for statistical data analysis using the BioStat software. Results: NKIRAS1 expression was downregulated in 75% of RCC samples (9 of 12) compared with surrounding normal tissue. High grade tumors (3 and 4) showed lower expression of NKIRAS1 at the mRNA level than tumors of low grade (1 and 2). No significant association was found between gene expression level and gender or age. Analysis of NKIRAS1 gene copy number was performed in 19 tumor samples. Changes in the copy number of NKIRAS1 gene were observed in 64% (9 of 14) of cRCC samples. 9 samples displayed ratio ( 0.85) and were considered as normal copy number. Changes in NKIRAS1 gene copy number were detected in all 3 benign oncocytomas, 1 papillary cancer and 1 sarcoma, where hemizygous deletion was observed. No changes in methylation status of NKIRAS1 were found in RCC. Conclusions: We have validated the results of NotI microarray analysis of NKIRAS1 gene in RCC. It was shown the decreased expression level of NKIRAS1 in this type of tumor. 2010-01-01T00:00:00Z http://dspace.nbuv.gov.ua:80/handle/123456789/138606 Comparative investigation of the effect of Ukrain on growth of ascite and solid forms of ehrlich’s carcinoma Susak, Ya.M.; Skivka, L.M.; Rudik, M.P.; Pozur, V.V.; Liubunya, A.V. Ukrain (NSC-631570) is a cytostatic and immunomodulating semisynthetic compound of thiophosphate-modified alkaloids of Chelidonium majus L. It has selective cytotoxicity against cancer cells without healthy cells damaging. Dosage range, methods of introduction and duration of administration of the drug vary. Aim: To carry out comparative investigation of effect of Ukrain on growth of different form of Ehrlich’s carcinoma. Methods: Ehrlich’s carcinoma cells were transplanted intraperitoneally and subcutaneously between the scapulas. Ukrain was administered intraperitoneally for 6 days (0.25 mg per mice of 20 g, it’s 0.1 LD50). The effect of drug on tumor growth was evaluated by the indexes of tumor growth inhibition (in the case of solid form), total number of tumor cells in ascites, number of viable tumor cells (in the case of ascitic form) and average life span of experimental animals. Cell cycle distribution of cancer cells was determined by flow cytometry. The number of circulating phag ocytes was determined by flow cytometry with use of FITC-labelled S. aureus Cowan. Results: Intraperitoneal Ukrain administration in mice with ascite form of Ehrlich’s carcinoma resulted in moderate tumor growth retardation, but was accompanied by acute local inflammation and caused reduction of life span of experimental animals. In mice bearing solid form of Ehrlich’s carcinoma treatment with Ukrain led to significant tumor growth inhibition and slight increase of life span. In mice bearing both of tumor variants treatment with drug caused restitution of the number of circulating phagocytes in peripheral blood, more expressed in mice bearing ascite tumor variant. Conclusion:Thus, anticancer activity of the Ukrain is more expressed in the case of solid variant of Ehrlich’s carcinoma. This effect is mediated by direct apoptotic action of drug on Ehrlich’s carcinoma cells and positive immunomodulating effect on tumor-bearing organism. 2010-01-01T00:00:00Z http://dspace.nbuv.gov.ua:80/handle/123456789/138605 Jurkat/A4 cells with multidrug resistance exhibit reduced sensitivity to quercetin Philchenkov, A.; Zavelevich, M.; Savinska, L.; Blokhin, D. Background: While multidrug resistance of cancer cells is a well-known phenomenon, little is known on the cross resistance between cytotoxic chemotherapeutical agents and unrelated substances such as natural flavonoids. Aim: To compare the effects of cytotoxic drug, vepeside and natural flavonoid, quercetin in Jurkat cells and their multidrug-resistant subline Jurkat/A4, in particular to analyze the effector mechanisms of apoptosis and the profiles of several pro- and antiapoptotic proteins in these cells upon exposure to vepeside or quercetin. Methods: Apoptosis and poly (ADP-ribose) polymerase cleavage were assessed by flow cytometry. Expression of apoptosisrelated proteins was analyzed by Western blotting. Results: Jurkat/A4 cells are less sensitive to antiproliferative effects of quercetin as compared with the parental Jurkat cell line. While vepeside as well as quercetin initially induces apoptosis in both cell lines, the following survival of the exposed cells is essentially different. In resistant Jurkat/A4 cells, vepeside or quercetin treatment activates significantly less caspase-9 and -3 as compared with that in the parental cells. The expression of Bad and BNip1 proteins in Jurkat/A4 cells is lower than in the parental cell line. At the same time, XIAP and CAS levels in Jurkat/A4 cells increase. Upon apoptosis induction, XIAP and CAS levels in Jurkat cells decrease, this effect being negligible in resistant cells. Conclusion: Multidrug-resistant Jurkat/A4 cells exhibit reduced sensitivity to cytotoxic effects of quercetin. The expression profile of Jurkat/A4 cells is characterized by the increased levels of XIAP and CAS representing the endogenous inhibitors of apoptosis. 2010-01-01T00:00:00Z http://dspace.nbuv.gov.ua:80/handle/123456789/138604 In vivo and In vitro effect of a nutrient mixture on human hepatocarcinoma cell line SK-HEP-1 Roomi, M.W.; Roomi, N.W.; Kalinovsky, T.; Niedzwiecki, A.; Rath, M. Long-term survival of patients with hepatocellular carcinoma (HCC), a common cancer worldwide, remains poor, due to metastasis and recurrence. Aim: To investigate the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract on human HCC cell line Sk-Hep-1 In vivo and In vitro. Methods: After one week of isolation, 5–6 week old male athymic nude mice were inoculated with 3 x 106 SK-Hep-1 cells subcutaneously and randomly divided into two groups; group A was fed a regular diet and group B a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighed and processed for histology. We also tested the effect of NM In vitro on SK-Hep-1 cells, measuring cell proliferation by MTT assay, invasion through Matrigel, apoptosis by green caspase detection kit, MMP secretion by zymography, and morphology by H&E staining. Results: NM inhibited tumor weight and burden of SK-Hep-1 xenografts by 42% and 33% respectively. In vitro, NM exhibited 33% toxicity over the control at 500 and 1000 μg/ml concentration. Zymography demonstrated MMP-2 and MMP-9 secretion which was inhibited by NM in a dose dependent fashion, with virtual total inhibition at 1000 μg/ml. Invasion through Matrigel was inhibited at 100, 500 and 1000 μg/ml by 53%, 83% and 100% respectively. NM induced slight apoptosis at 100 μg/ml, and profound apoptosis at 500 μg/ml and 1000 μg/ml concentration. Conclusions: These results suggest that NM has therapeutic potential in treatment of HCC. 2010-01-01T00:00:00Z