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EN
This work was undertaken to compare cytotoxicity, DNA damaging properties and effect on DNA cleavage by topoisomerase II of the anthracycline drug doxorubicin (DOX) and its two derivatives with a formamidino group containing a cyclic amine moiety such as morpholine (DOXM) or hexamethyleneimine (DOXH). The tetrazolium dye colorimetric assay was used to determine the cytotoxic activity of anthracyclines toward L1210 leukemia cells. DNA damage was measured by alkaline elution technique. The effect of anthracyclines on DNA cleavage was studied in a cell-free system containing supercoiled pBR322 DNA and purified human topoisomerase II. The cytotoxicity data and the results of studies on the mechanism of DNA break formation by anthracyclines at the cellular level and in the cell-free system showed that the presence of the formamidino group in the doxorubicin molecule reduced its ability to stimulate DNA cleavage by DNA topoisomerase II. Conclusion: DNA topoisomerase II is not a primary cellular target for DOXM or DOXH. An advantageous feature of formamidinoanthracyclines is their mechanism of cytotoxic action which is not related to the inhibition of DNA topoisomerase II. Therefore this class of anthracyclines seems to be a good source for selection of an anticancer drug directed toward cancer cells with the developed multidrug resistance attributed to the presence of altered DNA topoisomerase II.
EN
The aim of the study was to verify the thesis that the cardiotoxic effects of doxorubicin are connected with activation of the ubiquitin - proteasome pathway followed by protein degradation. The expression of myocardial damage markers - fatty acid binding protein (H-FABP) and brain natriuretic peptide (BNP) was evaluated in rat fetal cardiomyocytes simultaneously treated with doxorubicin and the proteasome inhibitor - bortezomib. The level of H-FABP and BNP protein under the influence of doxorubicin was decreased below the detection threshold with unchanged (H-FABP) or elevated (BNP) mRNA expression level. Against the expectations, the inhibitor of proteasome did not abolish this effect. The observed abnormal expression of BNP and H-FABP protein after doxorubicin treatment makes their diagnostic significance in anthracycline cardiotoxicity questionable.
EN
Anticancer antibiotics, anthracyclines, may cause severe left ventricle systolic dysfunction that is associated with poor clinical prognosis. In this review, we discuss the mechanisms and risk factors of anthracycline-induced cardiotoxicity and cardiomyopathy, among which the cumulative administered dose of anthracyclines and concurrent cardiovascular diseases are the most important. We also present strategies for primary and secondary prevention of anthracycline-induced cardiotoxicity via rigorous assessment of early signs of left ventricular dysfunction.
EN
Chitosan (Chi) is an extremely promising natural biopolymer with remarkable potency for the development of drug and vaccine delivery nanosystems. Various Chi derivatives are used to form nanoparticles (NPs) with unique properties. However, the efficacy of the therapy delivered by Chi NPs depends significantly on NP biodistribution in the body. The aim of this study was the analysis of biodistribution of NPs formed by succinoyl Chi and loaded with doxorubicin (SCNPDOX). We compared the distribution of free DOX and SCNP-DOX after intravenous (i.v.) and intranasal (i.n.) delivery into tumour-bearing mice. Distribution of DOX and SCNP-DOX was comparable after i.v. injection while they differed significantly after i.n. instillation.
EN
The study aimed to check the effectiveness of anticancer therapy combining a vascular-disruptive drug (combretastatin phosphate, CA4P) and a liposomal formulation of a chemotherapeutic (doxorubicin). CA4P was synthesized in our laboratory according to a previously described procedure. The antivascular drug and long-circulating doxorubicin-loaded liposomes were used to treat B16-F10 murine melanoma experimental tumors. Seventy-four hours after drug administration, a decrease in the number of tumor blood vessels was apparent and necrotic areas within tumors were visible. Combination therapy consisting of alternate administrations of CA4P and liposomal doxorubicin yielded greater inhibition of tumor growth than monotherapies alone. The best therapeutic results were obtained with the antivascular drug administered intratumorally every second day at 50 mg/kg body mass. In the case of combined therapy, the best results were obtained when the vascular-disruptive agent (CA4P) and the antineoplastic agent (liposomal doxorubicin) were administered in alternation.
EN
The results presented demonstrate the influence of aromatic indolinic aminoxyls: 1,2-dihydro-2-ethyl-2-phenyl-3H-indole-3-phenylimino-1-oxyl (IA-C2) and 1,2-dihydro-2-octadecyl-2-phenyl-3H-indole-3-phenylimino-1-oxyl (IA-C18) on oxidation of lipids and proteins of cardiac sarcoplasmic reticulum membranes. We have used doxorubicin and t-butyl hydroperoxide as agents inducing oxidative stress in isolated rat cardiac sarcoplasmic reticulum membrane system. Carbonyl groups were measured as the end product of membrane protein oxidation, and thiobarbituric acid reactive substances were assessed as a marker of lipid peroxidation. Inhibition of peroxidation of certain membrane components depends on the length of acyl chain. Aminoxyl IA-C2 inhibits the lipid peroxidation process while IA-C18 is an efficient protector against protein oxidation.
EN
Introduction: Sarcomatoid renal-cell carcinoma is a very rare cancer characterised with aggressive course of disease and poor prognosis. At present there are no standards of care for this histologic subtype of renal cell carcinoma resistant to various forms of systemic treatment. Methods: The study describes a case of 58 year old woman after left nephrectomy for clear cell carcinoma with sarcomatoid component and after resection of right-kidney tumour for synchronous clear cell carcinoma who received first-line bevacizumab and temsirolimus under the clinical trial, and then second-line chemotherapy based on gemcitabine and doxorubicin and ifosfamide-based third-line chemotherapy. The patient underwent pulmonary metastasectomy twice, and once a metastasectomy for liver metastases. Conclusions: Surgery (including metastases treatment) followed by the systemic chemotherapy seems to be correct option of treatment in patients with renal cell carcinoma with sarcomatoid features. The development of optimum method of systemic treatment requires further prospective randomised trials.
EN
Previously performed experiments showed that methylxanthines, especially caffeine, may protect cells against cytostatic or cytotoxic effects of several aromatic compounds. One of the proposed mechanisms of this protection is based on stacking interactions between π electron systems of polycyclic aromatic molecules. In this work, we demonstrate that caffeine and other methylxanthines - pentoxifylline and theophylline - significantly decrease mutagenicity of the anticancer aromatic drugs daunomycin, doxorubicin and mitoxantrone. The spectrophotometric titration of these aromatic compounds by methylxanthines indicated formation of mixed aggregates. The concentrations of free active forms of the drugs decreased when the concentrations of methylxanthines increased in the mixture. Therefore, likely methylxanthines may play a role of scavengers of the free active forms of daunomycin, doxorubicin and mitoxantrone.
EN
The main side effects of the administration of doxorubicin, a widely used anticancer drug, is the generation of a reactive oxygen species (ROS) in normal cells. As a result, redox disorders and secondary oxidative stress are developed. Doxorubicin ROS generation is attributed to enzymes that are produced abundantly in hepatocytes. Oxidative stress has been a well-known risk factor of doxorubicin-related toxicity. However, in addition, according to the data collected in the last decade, changes in thyroxin status can propagate ROS generation, and, thus, initiate the doxorubicin hepatic effect. Moreover, both compounds have an impact on the cell metabolism. The aim of the study was to verify the thesis that thyroxin can modulate the effect of doxorubicin with regard to redox status and lipid metabolism disorders. In our work, we determined the ratio of NADP+/ NADPH and NAD+/NADH in liver homogenates, blood ketone bodies and triglycerides in the liver and blood in rats treated with doxorubicin and thyroxin. Our results indicate that thyroxin has an insignificant effect on NAD+/NADH, NADP+/NADPH ratios and on hepatic and blood triglycerides. Moreover, thyroxin administration normalized the level of blood ketone bodies that was disturbed by doxorubicin.
EN
The redox cycle triggered by one electron reduction of doxorubicin and tirapazamine - both anticancer agents - leads to superoxide production. This superoxide production itself removes one iron atom from the [4Fe-4S] cluster, being an active center of aconitase. In addition, the incurred changes in cell redox equilibrium may affect lipid metabolism. The aim of the study was to evaluate a concomitant effect of both drugs on hepatic aconitase activity and triglycerides level. In our study, doxorubicin (1.8 mg/kg b.w.) was administered intraperitoneal (i.p.) six times, once a week, within male Wistar rats, to achieve a cumulative dose of 10.8 mg/kg b.w. Two hours before every doxorubicin administration, tirapazamine in the dose of either 5 or 10 mg/kg b.w. was also i.p. injected. A week after withdrawing drug administration, the liver was taken for biochemical analysis. Therein, an increase in aconitase activity and a decrease in triglycerides level was seen in all groups exposed to doxorubicin. Our work demonstrated that tirapazamine administration had no influence on both tested parameters, but its higher dose rate normalized aconitase activity affected by doxorubicin.
EN
The in vivo antioxidant properties of green tea extract (GTE) were investigated against doxorubicin (DOX) induced cardiotoxicity in rats. In this experiment, 48 Wistar albino rats (200ñ250 g) were divided into eight groups (n = 6). Control group received normal saline for 30 days. Cardiotoxicity was induced by DOX (20 mg/kg i.p.), once on 29th day of study and were treated with GTE (100, 200 and 400 mg/kg, p.o.) for 30 days. Aspartate aminotransferase (AST), creatinine kinase (CK), lactate dehydrogenase (LDH), lipid peroxidation (LPO), cytochrome P450 (CYP), blood glutathione, tissue glutathione, enzymatic and non-enzymatic antioxidants were evaluated along with histopathological studies. DOX treated rats showed a significant increased levels of AST, CK, LDH, LPO and CYP, which were restored by oral administration of GTE at doses 100, 200 and 400 mg/kg for 30 days. Moreover, GTE administration significantly increased the activities of glutathione peroxidase (GPX), glutathione reductase (GR), glutathione s-transferase (GST), superoxide dismutase (SOD) and catalase (CAT), in heart, which were reduced by DOX treatment. In this study, we have found that oral administration of GTE prevented DOX-induced cardiotoxicity by accelerating heart antioxidant defense mechanisms and down regulating the LPO levels to the normal levels.
EN
The significant progress in the treatment of non-Hodgkin lymphomas, translating into prolongation of overall survival results in the manifestation of long-term adverse events, like anthracycline-related cardiotoxicity. Despite the dose-dependent cardiac dysfunction and the presence of risk factors, the increasing probability of cardiotoxicity arises from individual predisposition. Identification of high- -risk patients gives the opportunity to implement the prevention strategies to reduce the incidence of cardiac complications. The study evaluated the utility of biomarkers: N-terminal B-type natriuretic peptide, troponin I and electrocardiography with spatial QRS-T angle assessment, as indicators of individual sensitivity of cardiomyocytes to doxorubicin resulting in myocardial damage. Thirty-five treatment-naïve patients at increased risk of cardiotoxicity, were subjected prospectively during (R)-CHOP treatment to echocardiographic assessment and analysis of biomarkers: TnI and NT-proBNP plasma level and spatial QRS-T assessment before and 24 hours after each cycle of chemotherapy. The analysis of QRS-T angle was consistent with the results of NT-proBNP assessment and allowed to identify, after the first cycle of chemotherapy, patients at increased risk of developing cardiovascular complications, who require thorough echocardiographic analysis and primary cardioprotection implementation. Our data did not reveal the role of TnI in the identification of cardiac events. Our findings, though promising, should be confirmed in a larger group of patients in real-life or clinical trials.
EN
STI571 (imatinib mesylate; Gleevec®) is an inhibitor that targets the tyrosine kinase activity of Bcr-Abl present in chronic myelogenous leukemia (CML) cells. Some preclinical studies have demonstrated that the combination of STI571 with chemotherapeutic drugs results in enhanced toxicity in Bcr-Abl-positive leukemias. We investigated the potential benefit of using STI571 to down-regulate Bcr-Abl activity for the enhancement of doxorubicin anti-proliferative action in K562 cell line derived from blast crisis of CML. At low concentrations of both drugs (40 nM doxorubicin combined with STI571 in the range of 100-150 nM), the antiproliferative effects were mainly due to cellular differentiation as assessed by benzidine staining for hemoglobin synthesis level and real-time PCR for γ-globin expression. Higher concentrations of STI571 used in combinations with doxorubicin caused mainly apoptosis as shown by DNA degradation and nuclear fragmentation visualized by fluorescence microscopy after DAPI staining, changes in cell morphology observed after Giemza-May Grünwald staining and cellular membrane organization estimated by flow cytometry after Annexin V staining. As compared with either drug alone, cotreatment with STI571 and DOX induced stronger cellular responses. A low concentration of STI571 in combination with a low concentration of DOX might be tested as an alternative approach to increasing the efficacy of chemotherapy against CML.
EN
Natural polyphenolic compounds produced by plant exhibit many pharmacological effects including antioxidant, chemopreventive as well as anticancer properties. This study was conducted to investigate the effect of cynarin ( from Artichoke, Cynara scolymus) and isoliquiritin (from Licorice, Glycyrrhiza uralensis) on doxorubicin (positive control) cytotoxicity in different cell lines including normal (Fibroblasts MCR-5 and Myoblasts H9c2) and cancer (colorectal HCT-116 and hepatocellular HEP-G2) cell lines. The cytotoxic effect of doxorubicin, isoliquiritin and cynarin alone or in different combination was studied on cancer cell lines as well as normal cell lines. The results obtained indicated that both cynarin and isoliquiritin enhance the cytotoxicity of doxorubicin. Both cynarin and isoliquiritin also reduce the cardiotoxicity of doxorubicin on normal cardiac cell lines. The combination of the three compounds (cynarin, isoliquiritin and doxorubicin) result in decrease the cytotoxicity of doxorubicin, which may indicate the presence of interaction and/or antagonism effect between cynarin and isoliquiritin. Cynarin was found to enhance the growth of (HCT-116 and HEP-G2) this might suggest avoiding use of Artichoke in subjects’ susceptibility for these cancers. All results were evaluated using statistical path and showed significant findings. The mechanism of enhanced doxorubicin’s cytotoxicity by cynarin or isoliquiritin also require further investigation to explain the increasing and/or the decreasing effect of these polyphenolic compounds on cytotoxicity of doxorubicin. The current finding can help to start with safe minimum dose of two or three combination of compounds in the context of clinical trials and practice.
EN
Neuroblastoma is the most common extra-cranial solid tumor of childhood and it is characterized by the presence of a glycosphingolipid, GD2 ganglioside. Monoclonal antibodies targeting the antigen are currently tested in clinical trials. Additionally, several research groups reported results revealing that ganglioside-specific antibodies can affect cellular signaling and cause direct cytotoxicity against tumor cells. To shed more light on gene expression signatures of tumor cells, we used microarrays to analyze changes of transcriptome in IMR-32 human neuroblastoma cell cultures treated with doxorubicin (DOX) or a mouse monoclonal antibody binding to GD2 ganglioside 14G2a (mAb) for 24 h. The obtained results highlight that disparate cellular pathways are regulated by doxorubicin and 14G2a. Next, we used RT-PCR to verify mRNA levels of selected DOX-responsive genes such as RPS27L, PPM1D, SESN1, CDKN1A, TNFSF10B, and 14G2a-responsive genes such as SVIL, JUN, RASSF6, TLX2, ID1. Then, we applied western blot and analyzed levels of RPS27L, PPM1D, sestrin 1 proteins after DOX-treatment. Additionally, we aimed to measure effects of doxorubicin and topotecan (TPT) and 14G2a on expression of a novel human NDUFAF2 gene encoding for mimitin protein (MYC-induced mitochondrial protein) and correlate it with expression of the MYCN gene. We showed that expression of both genes was concomitantly decreased in the 14G2a-treated IMR-32 cells after 24 h and 48 h. Our results extend knowledge on gene expression profiles after application of DOX and 14G2a in our model and reveal promising candidates for further research aimed at finding novel anti-neuroblastoma targets.
EN
The objective of this study was to demonstrate diff erences in the gene expression of human cervical cancer cells (HeLa) and vinblastine-resistant KB-V1 subline treated with doxorubicin alone and combination of Selol 5% and doxorubicin. Ongoing studies seek to clarify the mechanism of action of Selol in diff erent types of cancer cells, including those which show multidrug resistance. Cells treatment with the tested compounds in the group of genes tested in HeLa cells causes other changes than in KB-V1 cells. In the resistant cells, exposure to Selol 5% and doxorubicin, released the cytotoxic eff ects by changing the expression of ABCC2 and BCL2L1 genes. The observed dependence also allows better understanding the molecular mechanisms of resistance in the KB-V1 cell line.
PL
Celem pracy było wykazanie różnic w ekspresji genów komórek ludzkiego nowotworu szyjki macicy (HeLa) i opornej na winblastynę podlinii KB-V1, poddanych działaniu samej doksorubicyny oraz po łącznym podaniu Selolu 5% i doksorubicy. Prowadzone badania zmierzają do wyjaśnienia mechanizmu działania Selolu w różnych typach komórek nowotworowych, w tym opornych wielolekowo. Poddanie komórek działaniu testowanych związków powoduje inne zmiany w grupie badanych genów w komórkach HeLa niż w komórkach KB-V1. Łączne podanie Selolu 5% i doksorubicyny wyzwala efekt cytotoksyczny w komórkach opornych KB-V1, co przypuszczalnie jest związane ze zmianą ekspresji genów ABCC2 i BCL2L1. Zaobserwowana zależność pozwala także lepiej zrozumieć molekularne podłoże oporności komórek linii KB-V1.
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