Genotoxicity Of Doxorubicin Research Articles

Baicalin and Baicalein Enhance Cytotoxicity, Proapoptotic Activity, and Genotoxicity of Doxorubicin and Docetaxel in MCF-7 Breast Cancer Cells.

Breast cancer is a major health concern and the leading cause of death among women worldwide. Standard treatment often involves surgery, radiotherapy, and chemotherapy, but these come with side effects and limitations. Researchers are exploring natural compounds like baicalin and baicalein, derived from the Scutellaria baicalensis plant, as potential complementary therapies. This study investigated the effects of baicalin and baicalein on the cytotoxic, proapoptotic, and genotoxic activity of doxorubicin and docetaxel, commonly used chemotherapeutic drugs for breast cancer. The analysis included breast cancer cells (MCF-7) and human endothelial cells (HUVEC-ST), to assess potential effects on healthy tissues. We have found that baicalin and baicalein demonstrated cytotoxicity towards both cell lines, with more potent effects observed in baicalein. Both flavonoids, baicalin (167 µmol/L) and baicalein (95 µmol/L), synergistically enhanced the cytotoxic, proapoptotic, and genotoxic activity of doxorubicin and docetaxel in breast cancer cells. In comparison, their effects on endothelial cells were mixed and depended on concentration and time. The results suggest that baicalin and baicalein might be promising complementary agents to improve the efficacy of doxorubicin and docetaxel anticancer activity. However, further research is needed to validate their safety and efficacy in clinical trials.

Essential oil from Pelargonium graveolens shows anticancer activity and ameliorates the DNA damage and genotoxicity of doxorubicin in mice

Essential oil from Pelargonium graveolens shows anticancer activity and ameliorates the DNA damage and genotoxicity of doxorubicin in mice

In vitro hematotoxicity of Vernonanthura polyanthes leaf aqueous extract and its fractions

Vernonanthura polyanthes, popularly known as ‘assa-peixe’, is widely used in Brazil for therapeutic purpose mainly to treat respiratory tract problems. However, few studies investigated its chemical safety. In this way, we first obtained the V. polyanthes leaf aqueous extract (VpLAE) and three fractions (aqueous; n-butanol, n-BF; and ethyl acetate), and we chemically characterized this material. Then, the cytogenotoxic potential of the VpLAE and its fractions was investigated against human erythrocytes and lymphocytes using Trypan blue exclusion test of cell viability and CometChip. The phytochemical screening of V. polyanthes leaf revealed the presence of total phenolic compounds, flavonoids, tannins, coumarins, terpenic compounds, and cardioactive heterosides. n-BF presented the highest total phenolic, flavonoids, and tannins contents and, consequently, the highest antioxidant activity, according to the DPPH free radical scavenging method. Although the VpLAE and its fractions did not cause death of erythrocytes, the cells acquired an echinocytic form. Regarding lymphocytes, VpLAE and its fractions presented cytotoxicity and genotoxicity. When VpLAE or its fractions were co-treated with doxorubicin (DXR), a recognized cytotoxic drug, we observed an enhancement of DXR cytotoxicity against lymphocytes, but the DXR genotoxicity decreased around 15%. Since the VpLAE and its fractions increased the DXR cytotoxicity and decreased its genotoxicity, further studies should be conducted for the development of an adjuvant drug from this extract to reduce the side effects of chemotherapy. Moreover, the indiscriminate use of ‘assa-peixe’ by local people should be discouraged.

Combined study on clastogenic, aneugenic and apoptotic properties of doxorubicin in human cells in vitro

BackgroundDoxorubicin is a widely used anticancer drug due to its broad spectrum of antitumor activity. Various mechanisms have been proposed for its cytostatic activity, including DNA intercalation, topoisomerase II inhibition, generation of free radicals and apoptosis. The present study aims to further clarify the cytostatic activity of doxorubicin by its specific effect on (a) DNA damage, (b) micronucleation and (c) apoptosis, using a combination of different methods and cell systems such as human lymphocytes and HL-60 human leukemic cells. DNA lesions were analyzed by the alkaline comet assay in combination with formamidopyrimidine (Fpg) and human 8-oxoguanine (hOGG1) repair enzymes. Micronucleation was investigated by the Cytokinesis-Block Micronucleus assay (CBMN) in combination with Fluorescence In Situ Hybridization analysis. Impairment on mitotic apparatus was investigated by double immunofluorescence of β- and γ-tubulin. Apoptotic cell frequency was determined by the CBMN cytome assay. Complementary to the above, caspase-3 level was investigated by Western blot.ResultsIt was found that doxorubicin generates DNA breakage induced by oxidative damage in DNA bases, which can be repaired by the Fpg and hOGG1 enzymes. Increased micronucleus frequency was identified mainly through chromosome breakage and, at a lesser extent, through chromosome delay. Analysis of mitotic spindle showed disturbance of chromosome orientation and centrosome duplication and/or separation, leading to aneuploidy. Enhanced frequency of apoptotic leukemic cells was also observed. Caspase-3 seems to be involved in the generation of apoptosis.ConclusionsThe aforementioned findings derived from different treatment schedules, doses and time of exposure on primary versus transformed cells extend our knowledge about doxorubicin genotoxicity and contribute to the better understanding of the mechanisms by which doxorubicin induces genotoxic effects on human cells.

MODULATOR IMPACTS OF PROPOLIS EXTRACT AGAINST DOXORUBICIN MEDIATED CARCINOGENESIS ON HEPATOCELLULAR CARCINOMA AND Drosophila SOMATIC CELLS

The antitumor action of propolis is of clinical interest because of the need for new anticancer treatment agents. The present investigation intended to extract and assess the chemical content, cytotoxic action, the growth inhibitory activity and anticancer capability of Egyptian propolis versus Chinese propolis. This was carried out using water extract (WE) and ethanolic extract (EE) on the human hepatocellular carcinoma (HEp-2) cell line and the loss of heterozygosity (LOH) assay of Drosophila melanogaster somatic cells against the direct genotoxicity of doxorubicin. EPWE, EPEE, CPWE and CPEE extracts analyzed by HPLC showed that there were sensible and various concentrations of phenolic compounds in both. Total phenolics were determined to be 18.83, 34.87, 39.29 and 180.89 g-1 by using EPWE, CPWE, EPEE and CPEE extracts, respectively. Chinese propolis ethanol extract (CPEE) have major concentrations of total phenolics and phenolic acids and contained high concentrations of rutin (188.90 g/mL). The study of the antiproliferative capacity of propolis extractors against HEp-2 cancer cell lines showed that all the studied propolis extracts induce suppression of cell growth except CPWE extract; it gave 100% cell viability. The great majority of the propolis are strongly cytotoxic against HEp-2 cell line with 500 μg/ml CPEE. Also, PEE is the most effective in inhibition of HEp-2 cell proliferation compared to PWE. In Drosophila assay, treatment with propolis extract and DOX carcinogenic agent led to a reduction in the frequency of recombination compared to the treatment with DOX alone either in the post- and pre-treatments. In general, PEE exhibited powerful anti-proliferative effects than PWE. The ethanol extract provided the highest protection against Doxorubicin (DOR) induced genotoxicity, a fact that supports their anti-cancer activity. The results demonstrate that PEE is a good source of a natural antitumor operator able to inhibit cancer cell proliferation.

Reveal Cytotoxicity and Antigenotoxicity of Piper nigrum L. Ethanolic Extract and its Combination with Doxorubicin on CHO-K1 Cells

Black pepper (Piper nigrum L.), one of the most popular Indonesian spices has been reported to possess various therapeutic effects. The aim of this study is to evaluate the cytotoxicity and antigenotoxicity of black pepper ethanolic extract (BPE) and its combination with doxorubicin (Dox) on CHO-K1 cells. Based on thin layer chromatographyanalysis, BPE contained piperine.Under MTT assay, BPE showed cytotoxic effect with the IC50 value of 68 μg/mL and performed synergism in combination with Dox. In vitro micronucleus test using Giemsa staining revealed that BPE did not cause morphological changes qualitatively on CHO-K1 cells at concentration of 8.5 μg/mL, whereas using flow cytometry analysis showed that BPE could decrease the number of micronucleus (MN) formation induced by doxorubicin. In addition, BPE reduced the ROS level on the CHO-K1 cells which observed by reactive oxygen species (ROS) intracellular assay. The decrease in ROS level indicated that the antioxidant activity of BPE contribute to the antigenotoxicity. Furthermore, molecular docking performed that piperine interacted with DNA Topoisomerase II with docking score of -80.68. Overall,BPE performed cytotoxic effect in single treatment, increased the cytotoxicity and reduced the genotoxicity of doxorubicin. Thus, BPE has potential to be developed further as co-chemotherapeutic and antigenotoxic agent.Keywords: Cytotoxic, genotoxic, Piper nigrum L., CHO-K1, micronucleus

PH-Responsive Hybrid Organic-Inorganic Ruthenium Nanoparticles for Controlled Release of Doxorubicin

The current study aims at preparing biocompatible hybrid organic–inorganic ruthenium core–shell nanostructures (RuNPs) coated with polyvinylpyrrolidone (PVP) and polyoxyethylene stearate (POES). Thereafter, the core/shell RuNPs are loaded with doxorubicin (to form RuPDox) with a loading efficiency > 60%. RuPDox possesses exceptional stability and pH-responsive release kinetics with approx. 50% release of doxorubicin at up to 1 h exposure to an acidic endosomal environment. The cytotoxic effects of RuPDox are tested in vitro against breast cancer (MDA-MB-231), ovarian cancer (A2780), and neuroblastoma (UKF-NB-4) cells. Notably, although RuNPs have slight cytotoxicity only, RuPDox causes a synergistic enhancement of cytotoxicity when compared to free doxorubicin. Significant increase in free radicals formation, enhanced activity of executioner caspases 3/7, and higher expression of p53 and metallothionein is further identified due to the RuPDox treatment. Single-cell gel electrophoresis reveals no additional contribution of RuNPs to genotoxicity of doxorubicin. Moreover, RuPDox promotes significantly increased stability of doxorubicin in human plasma and pronounced hemocompatibility assayed on human red blood cells. The results imply a high potential of biocompatible hybrid RuNPs with PVP-POES shell as versatile nanoplatforms to enhance the efficiency of cancer treatment.

Vernonanthura polyanthes leaves aqueous extract enhances doxorubicin genotoxicity in somatic cells of Drosophila melanogaster and presents no antifungal activity against Candida spp.

Vernonanthura polyanthes (Spreng.) A.J. Vega & Dematt. (Asteraceae), known as "assa-peixe", has been used in ethnomedicine for the treatment of various diseases such as bronchitis, pneumonia, hemoptysis, persistent cough, internal abscesses, gastric and kidney stone pain. Moreover, some studies demonstrated that species of Genus Vernonia present antifungal activity. Due to the biological relevance of this species, the aim of this study was to investigate the toxic, genotoxic, antigenotoxic and antifungal potential of V. polyanthes leaves aqueous extract in somatic cells of Drosophila melanogaster or against Candida spp. The aqueous extract of the plant showed no toxic, genotoxic and antigenotoxic activity in the experimental conditions tested using the wing somatic mutation and recombination test (SMART/wing). However, when the extract was associated with doxorubicin, used in this work as a positive control, the mutagenic potential of doxorubicin was enhanced, increasing the number of mutations in D. melanogaster somatic cells. In the other hand, no inhibitory activity against Candida spp. was observed for V. polyanthes leaves aqueous extract using agar-well diffusion assay. More studies are necessary to reveal the components present in the V. polyanthes leaves aqueous extract that could contribute to potentiate the doxorubicin genotoxicity.

C(60) fullerene prevents genotoxic effects of doxorubicin in human lymphocytes in vitro

The self-ordering of C60 fullerene, doxorubicin and their mixture precipitated from aqueous solutions was investigated using atomic-force microscopy. The results suggest the complexation between the two compounds. The genotoxicity of doxorubicin in complex with C60 fullerene (С60+Dox) was evaluated in vitro with comet assay using human lymphocytes. The obtained results show that the C60 fullerene prevents the toxic effect of Dox in normal cells and, thus, С60+Dox complex might be proposed for biomedical application.

Influence of 3-aminobenzamide, an inhibitor of poly(ADP-ribose)polymerase, in the evaluation of the genotoxicity of doxorubicin, cyclophosphamide and zidovudine in female mice

Testing new chemical entities for genotoxicity is an integral part of the preclinical drug-development process. Lowering the detection limit and enhancing the sensitivity of genotoxicity assays is required, as the standard test-battery fails to detect some carcinogens (non-genotoxic) and weak genotoxins. One of the mechanisms that affect the detection of weak genotoxins is related with the DNA-repair efficiency of the cell system used. In the present study, 3-aminobenzamide (3-AB, 30mg/kg body-weight), a poly(ADP-ribose)polymerase inhibitor, was used to evaluate the DNA-damaging potential of zidovudine (AZT, 400mg/kgbw), doxorubicin (DOX, 5mg/kgbw) and cyclophosphamide (CP, 50mg/kgbw, as a positive control) and sucrose (SUC, 3g/kgbw, as a negative control) in Swiss female mice. The endpoints considered included micronucleus formation, DNA breakage (in peripheral blood lymphocytes, bone marrow and liver; comet assay) and chromosome aberrations, as well as immunohistochemistry of PARP-1 and phosphorylated histone H2AX (γ-H2AX). The results clearly indicate that the genotoxicity of zidovudine (AZT), doxorubicin (DOX) and cyclophosphamide (CP) was significantly increased in the combination treatments (3-AB+AZT, 3-AB+DOX, 3-AB+CP) as compared with the respective controls (treatment with AZT, DOX and CP alone). There was no increase in the genotoxicity per se after treatment with SUC, 3-AB or 3-AB+SUC, compared with the control (saline). Correlation analysis suggests that all genotoxicity parameters are well correlated with each other. The results clearly show that the genotoxicity of weak genotoxins can be enhanced and detected in the presence of 3-AB in mice. Thus, this approach can be used in the pre-clinical genotoxicity screening of weak genotoxins.

Assessment of the (anti)genotoxicity of brown propolis extracts from Brazilian Cerrado biome in a Drosophila melanogaster model

Among the great number of studies on the chemical and biological features of propolis from Brazil, none have been found on the evaluation of the biological properties of propolis from the Cerrado biome of Center-West Brazil. Propolis biological activities have principally been attributed to its phenolic compounds, notable for significant antioxidant properties responsible for reducing the toxic effects of free radicals, in addition to the anticancer potential of these compounds. The purposes of the present investigation were to determine the DPPH-radical-scavenging activities of extracts and fractions of a propolis sample from the Brazilian Cerrado biome and to evaluate their genotoxicities by performing the somatic mutation and recombination test (SMART) on wing cells of Drosophila melanogaster, using standard (ST) and high bioactivation (HB) crosses, with regular and enhanced levels of metabolizing enzymes of the cytochrome P450 type, respectively. The protective effects of propolis on the genotoxicity of doxorubicin (DXR), as well as the correlation between the radical-scavenging ability of propolis and its antigenotoxic potential were also evaluated. The ethanol extract of propolis (EEP) showed a significant level of phenolic compounds, as well as great antioxidant potential, unlike the hexane extract (HEP), of inexpressive radical-scavenging activity. Neither HEP nor EEP proved genotoxic to the offspring of ST and HB crosses, while co-treatments with DXR demonstrated the antigenotoxic properties of EEP on the ST cross, albeit proving genotoxic at the highest concentrations tested under conditions of high metabolic activation (HB cross). Solvent partitioning of EEP revealed expressive protective effects against DXR-induced genotoxicity exhibited by the n-butanol fraction, which were potentiated in HB strains, possibly due to the higher total phenolic content and more pronounced radical-scavenging properties of this fraction.

Genotoxicity of doxorubicin in F344 rats by combining the comet assay, flow‐cytometric peripheral blood micronucleus test, and pathway‐focused gene expression profiling

Doxorubicin (DOX) is an antineoplastic drug effective against many human malignancies. DOX's clinical efficacy is greatly limited because of severe cardiotoxicity. To evaluate if DOX is genotoxic in the heart, ~7-week-old, male F344 rats were administered intravenously 1, 2, and 3 mg/kg bw DOX at 0, 24, 48, and 69 hr and the Comet assays in heart, liver, kidney, and testis and micronucleus (MN) assay in the peripheral blood (PB) erythrocytes using flow cytometry were conducted. Rats were euthanized at 72 hr and PB was removed for the MN assay and single cells were isolated from multiple tissues for the Comet assays. None of the doses of DOX induced a significant DNA damage in any of the tissues examined by the alkaline Comet assay. Contrastingly, the glycosylase enzymes-modified Comet assay showed a significant dose dependent increase in the oxidative DNA damage in the cardiac tissue (P ≤ 0.05). In the liver, only the top dose induced significant increase in the oxidative DNA damage (P ≤ 0.05). The histopathology showed no severe cardiotoxicity but non-neoplastic lesions were present in both untreated and treated samples. A severe toxicity likely occurred in the bone marrow because no viable reticulocytes could be screened for the MN assay. Gene expression profiling of the heart tissues showed a significant alteration in the expression of 11 DNA damage and repair genes. These results suggest that DOX is genotoxic in the heart and the DNA damage may be induced primarily via the production of reactive oxygen species.

In vivo protective effect of Copaifera langsdorffii hydroalcoholic extract on micronuclei induction by doxorubicin

Copaifera lansdorffii Desf. is known as 'copaíba', 'copaiva' or 'paú-de-óleo', and is found in part of Brazil. The present study was undertaken to evaluate the genotoxic potential of C. langsdorffii leaf hydroalcoholic extract (CLE) and its influence on the genotoxicity induced by the chemotherapeutic agent doxorubicin (DXR) using the Swiss mouse peripheral blood micronucleus test. HPLC analysis of CLE using two monolithic columns linked in series allowed quantification of two major flavonoid heterosides, quercitrin and afzelin. Animals were treated with CLE by gavage at doses of 10, 20, 40 and 80 mg kg(-1) body weight per day, each for 20 days. Peripheral blood samples were collected at 24 and 48 h, and 7, 15 and 21 days after the beginning of the treatment. For the antigenotoxicity evaluation, the animals treated with different concentrations of CLE received DXR (15 mg kg(-1) body weight, intraperitoneal) at day 20. The peripheral blood samples were collected 24 and 48 h after the treatment with DXR. The results demonstrated that CLE itself was not genotoxic in the mouse micronucleus assay. In animals treated with CLE and DXR, the number of micronucleus was significantly decreased compared with animals receiving DXR alone. The putative antioxidant activity of one or more of the active compounds of CLE may explain the effect of this plant on DXR genotoxicity.

The effect of apigenin on cyclophosphamide and doxorubicin genotoxicity in vitro and in vivo

The aim of this study was to investigate the mutagenic and antigenotoxic effects of different doses of the flavonoid, apigenin, alone and in combination with the antitumor drugs, cyclophosphamide and doxorubicin, in vitro and in vivo. Using bacterial reverse mutation inhibition in vitro, with and without metabolic activation, the effect of apigenin (10 – 400 μg/plate) was studied on genotoxicity induced by cyclophosphamide (800 μg/plate) and by doxorubicin (0.2 μg/plate). Subsequent to a dose-finding study in vivo, CD1 mice were treated with either cyclophosphamide (40 mg/kg, i.p.) or doxorubicin (5 mg/kg, i.p.) with or without co-administration of apigenin (1–100 mg/kg, p.o.). Micronuclei were determined microscopically in blood smears and glutathione peroxidase (GPX), superoxide dismutase (SOD) and total antioxidative status (TAS) in whole blood, erythrocytes and plasma, respectively. Apigenin decreased doxorubicin-induced, but not cyclophosphamide-induced mutagenicity in vitro. In vivo, apigenin caused a statistically significant decrease in micronucleus frequency in response to cyclophosphamide, possibly due to active flavonoid metabolite formation or inhibition of cyclophosphamide metabolic activation. In animals treated with apigenin and doxorubicin, a significant decrease in micronucleus frequency was not observed, probably due to interindividual variability. No changes in GPX, SOD or TAS were observed in response to either cytotoxic agents or the flavonoid, possibly due to limited metabolic transformation of the drugs at the doses used. The results of the present study provide further evidence for the chemo-preventative properties of apigenin.

Protective effects of β-carotene against the genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster

beta-carotene (BC), pro-vitamin A, is an efficient antioxidant, effective in the neutralization of oxygen reactive species, which cause serious damage to DNA. Various studies have been conducted on the effectiveness of BC for chemoprevention of cancer and heart disease. Doxorubicin is a chemotherapeutic agent used for cancer treatment that generates free radicals. We examined the effects of BC (1, 2 and 4 mg/mL) on the genotoxicity of doxorubicin (0.125 mg/mL), using the wing spot test in Drosophila melanogaster (somatic mutation and recombination test). The BC alone had no significant effect on the frequency of mutant spots. However, it significantly reduced the number of spots caused by doxorubicin. We concluded that BC is not genotoxic and that it exerts protective effects against the genotoxic action of the chemotherapeutic free-radical generator doxorubicin.

Effect of organic tomato ( Lycopersicon esculentum ) extract on the genotoxicity of doxorubicin in the Drosophila wing spot test

The consumption of organic tomatoes (ORTs) reduces the risk of harmful effects to humans and the environment caused by exposure to toxic agrochemicals. In this study, we used the somatic mutation and recombination test (SMART) of wing spots in Drosophila melanogaster to evaluate the genotoxicity of ORT and the effect of cotreatment with ORT on the genotoxicity of Doxorubicin® (DXR, a cancer chemotherapeutic agent) that is mediated by free radical formation. Standard (ST) cross larvae were treated chronically with solutions containing 25%, 50% or 100% of an aqueous extract of ORT, in the absence and presence of DXR (0.125 mg/mL), and the number of mutant spots on the wings of emergent flies was counted. ORT alone was not genotoxic but enhanced the toxicity of DXR when administered concomitantly with DXR. The ORT-enhanced frequency of spots induced by DXR may have resulted from the interaction of ORT with the enzymatic systems that catalyze the metabolic detoxification of this drug.

Protection by Panax ginseng C.A. Meyer against the genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster

Panax ginseng is one of the most widely prescribed herbal medicines for the treatment of cancer, diabetes, chronic inflammation, and neurodegenerative and cardiovascular diseases. Since the use of alternative medicines in combination with conventional therapy may increase the risk of unwanted interactions, we investigated the possible genotoxicity of a water-soluble form of the dry root of P. ginseng (2.5, 5.0 or 10.0 mg/mL) and its ability to protect against the genotoxicity of doxorubicin (DOX; 0.125 mg/mL) by using the Drosophila melanogaster wing somatic mutation and recombination test (SMART) with standard and high-bioactivation crosses of flies. Panax ginseng was not genotoxic at the concentrations tested, whereas DOX-induced genotoxicity in marker-heterozygous flies resulted mainly from mitotic recombination. At low concentrations, P. ginseng had antirecombinogenic activity that was independent of the concentration of extract used. Recombination events may promote cancer, but little is known about the ability of P. ginseng to inhibit such recombination or modulate DNA repair mechanisms.

Protective effects of a mixture of antioxidant vitamins and minerals on the genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster

Antioxidant vitamins are able to deactivate highly bioactive molecules, such as free radicals, that are generated during cellular biochemical processes. Doxorubicin (DXR) is a cancer chemotherapeutic agent that generates free radicals as a byproduct. In the present study, the Drosophila melanogaster somatic wing spot test was used to evaluate the effects of a mixture of vitamins (Vitamins C, E, and beta-carotene) and minerals (copper, selenium, and zinc), commercially known as Vitergan Zinc Plus, on the genotoxicity of DXR in standard and high-bioactivation crosses of flies. 12.5, 25, and 50 mg/ml of the vitamin/mineral mixture by itself was nongenotoxic in the trans-heterozygous descendants of both crosses, while the mixture produced a significant reduction in the genotoxicity produced by 0.125 mg/ml DXR in the trans-heterozygous descendants of both crosses. The protective effect was observed when the larvae received either pre- or cotreatments of the multivitamin/mineral (MV) mixture. The results indicate that, under these experimental conditions, the MV mixture is not genotoxic; however, it protects against the genotoxic effects of the chemotherapeutic free-radical generator DXR.