Changes of c-myc expression in b16 melanoma cells induced by 8-chloroadenosine-3′, 5′-monophosphate and tiazofurin

The aim of this study was to investigate the in vitro effects of 8- chloroadenosine 3′, 5′-monophosphate (8-Cl-cAMP) and tiazofurin (TR) on the expression of c-myc gene in B16/F10 and B16/C3 mouse melanoma cells. Exponentially growing cells were treated with 8-Cl-cAMP or TR (5µmol - 25µmol) for 6h and 24h. The level of c-myc expression, estimated by RT-PCR, did not significantly change in B16/F10 cells after treatment with 8-Cl-cAMP or TR. Similar results were obtained in B16/C3 cells after treatment with 8-Cl-cAMP. The level of c-myc expression has shown a significant increase in B16/C3 cells after treatment with TR. Further studies of these agents will lead to better understanding of molecular mechanisms of their action.Physical chemistry 2004 : 7th international conference on fundamental and applied aspects of physical chemistry; Belgrade (Serbia); 21-23 September 200

Examination of Prooxidative Activity of Red Wine in Melanoma Cells

Melanoma is responsible for 75% of all deaths from skin cancer. Its lethality arises from its rapid progression, easy metastasis and drug-resistance as well. Red wine is a natural product rich in polyphenolic compounds with potent anticancer activities. It seems that in cancer cells these compounds behave as prooxidants initiating reactive oxygen species mediated cellular DNA breakage and consequent cell death. The aim of this study was to investigate prooxidative activity of red wine samples (Merlot variety, commercial as well as VCR1 and VCR101 clonal wines) in melanoma A375 cells, through measuring the relationship of reduced and oxidized form of glutathione (GSH/GSSG) and comparison with the GSH/GSSG ratio in control (untreated melanoma cells). The data obtained showed that tested red wine samples decrease GSH/GSSG ratio in A375 cells compared to control (4.6 ± 0), with the largest decrease noticed in treatment with VCR101 wine (0.66 ± 0.05)

Inactivation of melanoma cells irradiated with gamma rays and low energy protons

13th Balkan biochemical Biophysical Days and Meeting on Metabolic Disorders, October 12.-15., 2003, Kusadasi, Turke

Citotoksično dejstvo crvenog vina na tumorske ćelije u in vitro uslovima

Benefitni efekat crvenog vina na zdravlje ljudi od davnina je poznat. Među alkoholnim pićima crveno vino pokazuje najjači efekat protiv pojave oboljenja povezanih sa oksidativnim stresom, kao što su kardiovaskularna, neurodegenerativna oboljenja, dijabetes i kancer.1 U okviru ove studije ispitan je citotoksični efekat crvenog vina sorte merlo (komercijalno dostupno vino i vina dobijena od klonova VCR1 i VCR101) poreklom iz Crne Gore, berba 2011, u odnosu na ćelijsku liniju kancera pankreasa (PANC-1) i ćelijsku liniju melanoma (A375). Citotoksični efekat vina je određen SRB metodom kroz praćenje ćelijskog preživljavanja kancerskih ćelija 48 sati nakon tretmana sa tri različita zapreminska procenta analiziranih vina (5, 10 i 20%). Dobijeni rezultati su pokazali citotoksični efekat svih analiziranih vina na obe ćelijske linije. Ćelijsko preživljavanje nakon tretmana vinima se kretalo od 36 do 64%. Citotoksični efekat svih vina u odnosu na A375 ćelije je bio veći nego na PANC-1 ćelijsku liniju, što je od posebnog značaja ako se uzme u obzir rezistentnost melanomskih ćelija na postojeće terapeutske tretmane. Takođe, klonska sortna vina su pokazala veći citotoksični efekat na A375 ćelijsku liniju od komercijalno dostupnog vina. Dobijeni rezultati su ukazali da umereno konzumiranje crvenog vina sorte merlo sa specifičnim geografskim poreklom predstavlja dobar izvor bioaktivnih komponenata sa pozitivnim biološkim efektom, što je od posebne važnosti za klonska sortna vina.3. kongres biologa Srbije : osnovna i primenjena istraživanja, metodika nastave, 21-25. septembar 2022., Zlatibor, Srbij

Radiosensitivity of human ovarian carcinoma and melanoma cells to gamma-rays and protons

Introduction: Proton radiation offers physical advantages over conventional radiation. Radiosensitivity of human 59M ovarian cancer and HTB140 melanoma cells was investigated after exposure to gamma-rays and protons. Material and methods: Irradiations were performed in the middle of a 62 MeV therapeutic proton spread out Bragg peak with doses ranging from 2 to 16 Gy. The mean energy of protons was 34.88+/-2.15 MeV, corresponding to the linear energy transfer of 4.7+/-0.2 keV/mu m. Irradiations with gamma-rays were performed using the same doses. Viability, proliferation and survival were assessed 7 days after both types of irradiation while analyses of cell cycle and apoptosis were performed 48 h after irradiation. Results: Results showed that gamma-rays and protons reduced the number of viable cells for both cell lines, with stronger inactivation achieved after irradiation with protons. Surviving fractions for 59M were 0.91+/-0.01 for gamma-rays and 0.81+/-0.01 for protons, while those for HTB140 cells were 0.93+/-0.01 for gamma-rays and 0.86+/-0.01 for protons. Relative biological effectiveness of protons, being 2.47+/-0.22 for 59M and 2.08+/-0.36 for HTB140, indicated that protons provoked better cell elimination than gamma-rays. After proton irradiation proliferation capacity of the two cell lines was slightly higher as compared to gamma-rays. Proliferation was higher for 59M than for HTB140 cells after both types of irradiation. Induction of apoptosis and G2 arrest detected after proton irradiation were more prominent in 59M cells. Conclusions: The obtained results suggest that protons exert better antitumour effects on ovarian carcinoma and melanoma cells than gamma-rays. The dissimilar response of these cells to radiation is related to their different features

Radiation dose determines the method for quantification of DNA double strand breaks

Ionizing radiation induces DNA double strand breaks (DSBs) that trigger phosphorylation of the histone protein H2AX (gamma H2AX). Immunofluorescent staining visualizes formation of gamma H2AX foci, allowing their quantification. This method, as opposed to Western blot assay and Flow cytometry, provides more accurate analysis, by showing exact position and intensity of fluorescent signal in each single cell. In practice there are problems in quantification of gamma H2AX. This paper is based on two issues: the determination of which technique should be applied concerning the radiation dose, and how to analyze fluorescent microscopy images obtained by different microscopes. HTB140 melanoma cells were exposed to gamma-rays, in the dose range from 1 to 16 Gy. Radiation effects on the DNA level were analyzed at different time intervals after irradiation by Western blot analysis and immunofluorescence microscopy. Immunochemically stained cells were visualized with two types of microscopes: AxioVision (Zeiss, Germany) microscope, comprising an ApoTome software, and AxioImagerA1 microscope (Zeiss, Germany). Obtained results show that the level of gamma H2AX is time and dose dependent. Immunofluorescence microscopy provided better detection of DSBs for lower irradiation doses, while Western blot analysis was more reliable for higher irradiation doses. AxioVision microscope containing ApoTome software was more suitable for the detection of gamma H2AX foci

Carbon ions of different linear energy transfer (LET) values induce apoptosis and G2 cell cycle arrest in radio-resistant melanoma cells

Background and objectives: The main goal when treating malignancies with radiation is to deprive tumour cells of their reproductive potential. One approach is to induce tumour cell apoptosis. This study was conducted to evaluate the ability of carbon ions (C-12) to induce apoptosis and cell cycle arrest in human HTB140 melanoma cells. Methods: In this in vitro study, human melanoma HTB140 cells were irradiated with the 62 MeV/n carbon (C-12) ion beam, having two different linear energy transfer (LET) values: 197 and 382 keV/mu m. The dose range was 2 to 16 Gy. Cell viability was estimated by the sulforhodamine B assay seven days after irradiation. The cell cycle and apoptosis were evaluated 48 h after irradiation using flow cytometry. At the same time point, protein and gene expression of apoptotic regulators were estimated using the Western blot and q-PCR methods, respectively. Results: Cell viability experiments indicated strong anti-tumour effects of C-12 ions. The analysis of cell cycle showed that C-12 ions blocked HTB140 cells in G2 phase and induced the dose dependent increase of apoptosis. The maximum value of 21.8 per cent was attained after irradiation with LET of 197 keV/mu m at the dose level of 16 Gy. Pro-apoptotic effects of C-12 ions were confirmed by changes of key apoptotic molecules: the p53, Bax, Bcl-2, poly ADP ribose polymerase (PARP) as well as nuclear factor kappa B (NF kappa B). At the level of protein expression, the results indicated significant increases of p53, NF kappa B and Bax/Bcl-2 ratio and PARP cleavage. The Bax/Bcl-2 mRNA ratio was also increased, while no change was detected in the level of NF kappa B mRNA. Interpretation and conclusions: The present results indicated that anti-tumour effects of C-12 ions in human melanoma HTB140 cells were accomplished through induction of the mitochondrial apoptotic pathway as well as G2 arrest

Carbon quantum dots/silver based metal organic framework composites in light enhanced wound healing

In recent years researchers have developed new strategies to enhance the effectiveness of wound healing by combining nanoparticles and infra red (IR) light. For example, some studies have shown that nanoparticles can be used to enhance the absorption of near-infrared laser (NIR) light by tissues, leading to increased healing rates [1]. The influence of NIR light on proliferation, collagen production, and wound healing was tested on: keratocytes (HaCaT) and fibroblasts (MRC-5) cells that are used as model systems of human skin equivalents that comprise an epidermal and a dermal compartment of skin. Also, these cells were treated with carbon quantum dots/silver-based metal-organic framework composites (Ag-MoFs-NCDs and Ag-MoFs-SCDs), which previously showed high antibacterial activity [2], without and with laser light. Firstly, we have found the most convenient and effective CW laser intensity (16 mW/cm2) and illumination time (3 minutes), which is not too high and short enough to influence human cells' proliferation and metabolism positively. Additional chemical treatment with Ag-MoFs-NCDs and Ag-MoFs-SCDs results in a further increase in human cell viability. Our measurements showed that the proliferation index in laser-illuminated cells and cells treated with Ag-MoFs-SCDs was at the level of the untreated control. Furthermore, Ag-MoFs-SCDs treatment and laser illumination induced a mild, insignificant increase in cellular proliferation. On the other hand, Ag-MoFs-NCDs treatment led to a more pronounced, albeit not significant increase, in cellular proliferation, while Ag-MoFs-NCDs treatment combined with laser illumination significantly increased proliferation. Also, we have detected a mild change in collagen level estimated by hydroxyproline assay, which may indicate a positive outcome of combined laser illumination and treatment, taking into account that after 48 hours, a change in cell's response to the treatment could be noticed. Finally, based on migration assay, we observe a complete wound closure after 48 hours in fibroblast cells treated with Ag-MoFs-NCDs and near-infrared laser light, Fig. 1.IX International School and Conference on Photonics : PHOTONICA2023 : book of abstracts; August 28 - September 1, 2023; Belgrad

Anti-cancer and imaging potential of fluorescent black carrot Carbon Dot nanoparticles

Carbon Dots (CDs) are biocompatible, fluorescent, water-soluble, and stable nanoparticles with a high potential to be used for vast biomedical applications [1,2]. We explore the application of CDs produced from natural sources, black carrots, as anti-cancer and imaging agents. These nanoparticles suppress cell growth of three different cancer cell lines, cervical (HeLa), pancreatic (PANC-1), and melanoma (A375) cell lines in vitro. However, the cytotoxic effect against A375 cells stands out, with only 20% of viable cells left after treatment (Fig.1(a)), antimetastatic potential, and a selectivity index higher than two, which indicates that the efficacy against melanoma cells is significantly greater than the toxicity against non-malignant cells (MRC-5). Furthermore, after the cellular uptake, green fluorescence was visible in the cytosol of A375 cells (Fig. 1 (b)). On the other hand, the DAPI stain for DNA was visible as a blue light in the cell nucleus. Moreover, cells with a higher intensity of green fluorescence in the nucleus, Fig. 1 (c) indicated with arrows, were the cells with condensed chromatin in the mitotic phase of the cell cycle (Fig. 1 (d) and (e)), which indicates that CDs interact with chromatin and that they could be used as a marker of cells mitosis and proliferation. In summary, we have demonstrated the great anti-cancer potential of black carrot CDs, for image-guided anti-cancer therapy of melanoma that can be used to recognize cell proliferation.IX International School and Conference on Photonics : PHOTONICA2023 : book of abstracts; August 28 - September 1, 2023; Belgrad

The presence of acrylamide in various type of food products from the Serbian market

Acrylamide forms when some foods are prepared at temperatures usually above 120°C and in low moisture conditions, due to a Maillard reaction between certain amino acids, such as asparagine, and reducing sugars. Acrylamide is carcinogenic to experimental mice and rats, neurotoxic and probably also carcinogenic and genotoxic for humans. The aim of this study was to determine the presence of acrylamide in various groups of food products in which its formation is expected to occur during the production process. In the period December 2017 to March 2021, 529 samples of different types of food products were tested. Samples were collected from the Serbian market. Most of the tested foods, almost half of them (44%), were various types of biscuits. The presence of acrylamide was determined using LC-MS/MS accredited method, with a limit of quantification (LOQ) of 50 µg kg-1and a limit of detection (LOD) of 25 µg kg-1. All samples from the snack product and waffle product groups contained acrylamide. Acrylamide was detected in almost all (98.98%) fine bakery products and biscuits (90.43%). In contrast, only 15.38% of bakery products contained acrylamide. Most of the tested foods contained acrylamide, 83.74% of them.61st International Meat Industry Conference 26-29 September 2021, Zlatibor, Serbi