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BACKGROUND: Gastric cancer is the second leading cause of cancer-related death globally. Unfortunately, the survival rate of the gastric cancer patients who underwent chemotherapy following surgery has been less than a half. Besides, chemotherapy has many side effects. Current evidence suggests that some antidepressants like duloxetine have growth-inhibiting effects against a number of cancer cell lines. OBJECTIVE: Thus, the aim of this study was to determine the cytotoxic and genotoxic effects of duloxetine on gastric cancer. METHODS: In this regard, the cytotoxicity and genotoxicity of duloxetine were investigated in MKN45 and NIH3T3 cell lines by MTT assay and on peripheral blood lymphocytes by MN assay. For this purpose, cells were cultured in 96 wells plate. Stock solutions of duloxetine and cisplatin were prepared. After cell incubation with different concentrations of duloxetine (1, 10, 25, 50, 100 and 200 µL), MTT solution was added. For micronucleus assay fresh blood was added to RPMI culture medium 1640 supplemented, and different concentrations of duloxetine (1, 10, 25, 50, 100 and 200 µL) were added. RESULTS: The cytotoxicity of duloxetine on MKN45 cancer cell line and NIH3T3 normal cell line were studied followed by MTT assay. duloxetine exhibited higher IC50 in the MKN45 cells in comparison with the NIH3T3 cells. In addition, genotoxic effect of duloxetine was evaluated by micronucleus assay. The results revealed that duloxetine induced more DNA damage at 100 and 200 µM and no significant difference at 200 µM with respect to cisplatin, but it had less genotoxic effects at 100 and 50 µM concentrations. CONCLUSION: Although, in this study, duloxetine had less genotoxicity than cisplatin in concentrations under 200 µM and showed cytotoxic effects as well, due to its IC50, it cannot be considered as a better choice for gastric cancer therapies with respect to cisplatin as a common anticancer drug.

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ABSTRACT BACKGROUND: Gastric cancer is the second leading cause of cancer-related death globally. Unfortunately, the survival rate of the gastric cancer patients who underwent chemotherapy following surgery has been less than a half. Besides, chemotherapy has many side effects. Current evidence suggests that some antidepressants like duloxetine have growth-inhibiting effects against a number of cancer cell lines. OBJECTIVE: Thus, the aim of this study was to determine the cytotoxic and genotoxic effects of duloxetine on gastric cancer. METHODS: In this regard, the cytotoxicity and genotoxicity of duloxetine were investigated in MKN45 and NIH3T3 cell lines by MTT assay and on peripheral blood lymphocytes by MN assay. For this purpose, cells were cultured in 96 wells plate. Stock solutions of duloxetine and cisplatin were prepared. After cell incubation with different concentrations of duloxetine (1, 10, 25, 50, 100 and 200 μL), MTT solution was added. For micronucleus assay fresh blood was added to RPMI culture medium 1640 supplemented, and different concentrations of duloxetine (1, 10, 25, 50, 100 and 200 μL) were added. RESULTS: The cytotoxicity of duloxetine on MKN45 cancer cell line and NIH3T3 normal cell line were studied followed by MTT assay. duloxetine exhibited higher IC50 in the MKN45 cells in comparison with the NIH3T3 cells. In addition, genotoxic effect of duloxetine was evaluated by micronucleus assay. The results revealed that duloxetine induced more DNA damage at 100 and 200 μM and no significant difference at 200 μM with respect to cisplatin, but it had less genotoxic effects at 100 and 50 μM concentrations. CONCLUSION: Although, in this study, duloxetine had less genotoxicity than cisplatin in concentrations under 200 μM and showed cytotoxic effects as well, due to its IC50, it cannot be considered as a better choice for gastric cancer therapies with respect to cisplatin as a common anticancer drug.

RESUMO CONTEXTO: O câncer gástrico é a segunda principal causa de morte relacionada ao câncer globalmente. Infelizmente, a taxa de sobrevivência dos pacientes com câncer gástrico que se submeteram à quimioterapia após a cirurgia, tem sido inferior à metade. Além disso, a quimioterapia tem muitos efeitos colaterais. Evidências atuais sugerem que alguns antidepressivos como a duloxetina têm efeitos inibidores de crescimento contra um número de linhas de células cancerosas. OBJETIVO: Assim, o objetivo deste estudo foi determinar os efeitos citotóxicos e genotóxicos da duloxetina sobre o câncer gástrico. MÉTODOS: A este respeito, a citotoxicidade e a genotoxicidade da duloxetina foram investigadas em linhas celulares MKN45 e NIH3T3 por ensaio de MTT e por ensaio de MN em linfócitos periféricos de sangue. Para este efeito, as células foram cultivadas em 96 placas. Soluções de estoque de duloxetina e cisplatina foram preparadas. Após incubação celular com diferentes concentrações de duloxetina (1, 10, 25, 50, 100 e 200 μL), a solução de MTT foi adicionada. Para o teste do micronúcleo o sangue fresco foi adicionado ao meio de cultura RPMI 1640 suplementado, e as concentrações diferentes de duloxetina (1, 10, 25, 50, 100 e 200 μL) foram adicionadas. RESULTADOS: A citotoxicidade da duloxetina na linha celular cancerosa MKN45 e NIH3T3 linha celular normal foram estudadas e seguidas pelo ensaio de MTT. A duloxetina exibiu maior IC50 nas células MKN45 em comparação com as células NIH3T3. Além disso, o efeito genotóxico da duloxetina foi avaliado pelo ensaio de micronúcleos. Os resultados revelaram que a duloxetina induziu mais dano de DNA em 100 e 200 μM e não houve diferença significativa em 200 μM em relação à cisplatina, mas teve menos efeitos genotóxicos nas concentrações de 100 e 50 μM. CONCLUSÃO: Embora, neste estudo, a duloxetina tenha menos genotoxicidade do que a cisplatina em concentrações inferiores a 200 μm e também tenha mostrado efeitos citotóxicos, devido ao seu IC50, não pode ser considerada como uma escolha terapêutica melhor para o câncer gástrico no que diz respeito à cisplatina como uma droga anticâncer comum.

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BACKGROUND: Gastric cancer is known as the fourth most common cancer. Current treatments for cancer have damaged the sensitive tissues of the healthy body, and in many cases, cancer will be recurrent. Therefore, need for treatments that are more effective is well felt. Researchers have recently shifted their attention towards antipsychotic dopamine antagonists to treat cancer. The anticancer activities of aripiprazole remain unknown. OBJECTIVE: This study aimed to evaluate the efficacy and safety of aripiprazole on gastric cancer and normal cell lines. METHODS: In this regard, the cytotoxicity and genotoxicity of aripiprazole were investigated in MKN45 and NIH3T3 cell lines by methyl tetrazolium assay and on peripheral blood lymphocytes by micronucleus assay. For this purpose, cells were cultured in 96 wells plate. Stock solutions of aripiprazole and cisplatin were prepared. After cell incubation with different concentrations of aripiprazole (1, 10, 25, 50, 100 and 200 µL), methyl tetrazolium solution was added. For micronucleus assay fresh blood was added to RPMI culture medium 1640 supplemented, and different concentrations of aripiprazole (50, 100 and 200 µL) were added. RESULTS: The finding of present study showed that the IC50 of aripiprazole in the cancer cell line (21.36 µg/mL) was lower than that in the normal cell line (54.17 µg/mL). Moreover, the micronucleus assay showed that the frequency of micronuclei of aripiprazole at concentrations below 200 µM was much less than cisplatin. CONCLUSION: Aripiprazole can be a good cytotoxic compound and good candidate for further studies of cancer therapy.

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ABSTRACT BACKGROUND: Gastric cancer is known as the fourth most common cancer. Current treatments for cancer have damaged the sensitive tissues of the healthy body, and in many cases, cancer will be recurrent. Therefore, need for treatments that are more effective is well felt. Researchers have recently shifted their attention towards antipsychotic dopamine antagonists to treat cancer. The anticancer activities of aripiprazole remain unknown. OBJECTIVE: This study aimed to evaluate the efficacy and safety of aripiprazole on gastric cancer and normal cell lines. METHODS: In this regard, the cytotoxicity and genotoxicity of aripiprazole were investigated in MKN45 and NIH3T3 cell lines by methyl tetrazolium assay and on peripheral blood lymphocytes by micronucleus assay. For this purpose, cells were cultured in 96 wells plate. Stock solutions of aripiprazole and cisplatin were prepared. After cell incubation with different concentrations of aripiprazole (1, 10, 25, 50, 100 and 200 μL), methyl tetrazolium solution was added. For micronucleus assay fresh blood was added to RPMI culture medium 1640 supplemented, and different concentrations of aripiprazole (50, 100 and 200 μL) were added. RESULTS: The finding of present study showed that the IC50 of aripiprazole in the cancer cell line (21.36 μg/mL) was lower than that in the normal cell line (54.17 μg/mL). Moreover, the micronucleus assay showed that the frequency of micronuclei of aripiprazole at concentrations below 200 μM was much less than cisplatin. CONCLUSION: Aripiprazole can be a good cytotoxic compound and good candidate for further studies of cancer therapy.

RESUMO CONTEXTO: O câncer gástrico é conhecido como o quarto câncer mais comum. Os tratamentos atuais para o câncer danificaram os tecidos sensíveis do corpo saudável e, em muitos casos, o cancro será recorrente. Portanto, a necessidade de tratamentos que são mais eficazes é desejada. Recentemente, os pesquisadores mudaram sua atenção para os antagonistas antipsicóticos da dopamina para tratar o câncer. As atividades anticâncer de aripiprazol permanecem desconhecidas. OBJETIVO: Este estudo objetivou avaliar a eficácia e a segurança do aripiprazol no câncer gástrico e nas linhagens celulares normais. MÉTODOS: A este respeito, a citotoxicidade e a genotoxicidade do aripiprazol foram investigadas em linhas celulares MKN45 e NIH3T3 por ensaio de metil tetrazólio e em linfócitos periféricos de sangue por ensaio de micronúcleos. Para este efeito, as células foram cultivadas em 96 placas. As soluções de estoque de aripiprazol e cisplatina foram preparadas. Após incubação celular com diferentes concentrações de aripiprazol (1, 10, 25, 50, 100 e 200 μL), a solução de metil tetrazólio foi adicionada. Para o ensaio do micronúcleo o sangue fresco foi adicionado ao meio de cultura RPMI 1640 suplementado, e as concentrações diferentes de aripiprazole (50, 100 e 200 μL) foram adicionadas. RESULTADOS: O presente estudo mostrou que o IC50 de aripiprazol na linhagem celular cancerosa (21,36 μg/mL) foi menor do que na linha celular normal (54,17 μg/ mL). Além disso, o ensaio de micronúcleos demonstrou que a frequência de micronúcleos de aripiprazol em concentrações inferiores a 200 μM foi muito inferior à cisplatina. CONCLUSÃO: O aripiprazol pode ser um bom composto citotóxico e bom candidato para estudos adicionais da terapia do câncer.

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OBJECTIVE: Relacin is a synthetic molecule that targets RelA, an essential protein in a conserved bacterial stress response system. It was shown to inhibit bacterial growth. The aims of this study were to evaluate the antimicrobial effect of relacin combined with sodium hypochlorite (NaOCl) on Enterococcus faecalis biofilms and to evaluate the cytotoxicity of relacin. MATERIAL AND METHODS: 48-h E. faecalis OG1RF biofilms were treated by various concentrations of relacin in order to determine its inhibitory concentration. Then, the 48-h biofilms were treated either with 1-min NaOCl (0.01%, 0.05%) alone, or in combination of relacin. As a means of comparison, the biofilms of ΔrelA were also treated by 1-min NaOCl (0.01%, 0.05%, 0.25%). The treatment efficacy was determined by agar plate count assays. The cytotoxicity of relacin was examined on human gingival epithelial cells Ca9-22 and murine fibroblasts NIH-3T3 by a methyl thiazolyltetrazolium (MTT) assay and a lactate dehydrogenase assay. Statistical analysis was performed by one-way or two-way analysis of variance (ANOVA) with Bonferroni's post-hoc test and an independent Student's t-test. A significance level of p<0.05 was used. RESULTS: Relacin inhibited the growth of OG1RF biofilms partially at 8 mM and fully at 14 mM. The relacin (14 mM) and NaOCl combined treatment resulted in significantly higher treatment efficacy than NaOCl treatment alone. At 0.05% NaOCl, the combined treatment resulted in 5.65 (±0.19) log reduction in biofilm viability. The ΔrelA biofilms were more susceptible to NaOCl treatment than the wild type biofilms at 0.25% NaOCl. Relacin at 14 mM was not toxic to host epithelial cells and fibroblasts. CONCLUSIONS: The combination of relacin with a low concentration of NaOCl was effective and not cytotoxic.

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Abstract Objective Relacin is a synthetic molecule that targets RelA, an essential protein in a conserved bacterial stress response system. It was shown to inhibit bacterial growth. The aims of this study were to evaluate the antimicrobial effect of relacin combined with sodium hypochlorite (NaOCl) on Enterococcus faecalis biofilms and to evaluate the cytotoxicity of relacin. Material and Methods 48-h E. faecalis OG1RF biofilms were treated by various concentrations of relacin in order to determine its inhibitory concentration. Then, the 48-h biofilms were treated either with 1-min NaOCl (0.01%, 0.05%) alone, or in combination of relacin. As a means of comparison, the biofilms of ΔrelA were also treated by 1-min NaOCl (0.01%, 0.05%, 0.25%). The treatment efficacy was determined by agar plate count assays. The cytotoxicity of relacin was examined on human gingival epithelial cells Ca9-22 and murine fibroblasts NIH-3T3 by a methyl thiazolyltetrazolium (MTT) assay and a lactate dehydrogenase assay. Statistical analysis was performed by one-way or two-way analysis of variance (ANOVA) with Bonferroni's post-hoc test and an independent Student's t-test. A significance level of p<0.05 was used. Results Relacin inhibited the growth of OG1RF biofilms partially at 8 mM and fully at 14 mM. The relacin (14 mM) and NaOCl combined treatment resulted in significantly higher treatment efficacy than NaOCl treatment alone. At 0.05% NaOCl, the combined treatment resulted in 5.65 (±0.19) log reduction in biofilm viability. The ΔrelA biofilms were more susceptible to NaOCl treatment than the wild type biofilms at 0.25% NaOCl. Relacin at 14 mM was not toxic to host epithelial cells and fibroblasts. Conclusions The combination of relacin with a low concentration of NaOCl was effective and not cytotoxic.

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A espécie vegetal Qualea grandiflora (QG), popularmente conhecida como "pauferro", "pau-terra-da-folha-grande", "pau-terra" ou "pau-de-tucano", muito comum no Cerrado brasileiro, é bem conhecida devido às suas variadas propriedades terapêuticas. Suas indicações incluem ações preventivas no aparecimento de lesões de mucosa gástrica, efeitos analgésicos, antibacterianos, anti-inflamatórios e antifúngicos. Assim, os componentes da QG poderiam ter alguma ação sobre moléculas amplamente envolvidas em processos angiogênicos e de desenvolvimento/reparo, como a Metaloproteinase de matriz 14 (MMP-14) e o Fator Induzido por hipóxia 1α (HIF-1alfa). Dessa maneira, o objetivo deste estudo foi investigar os efeitos do extrato hidroalcoólico das folhas de QG na viabilidade celular e expressão de MMP-14 e HIF-1alpha em culturas de fibroblastos da linhagem NIH/3T3 e pré-osteoblastos da linhagem MC3T3-E1. Para o teste de viabilidade celular e expressão das moléculas, concentrações de 0.1; 1.0 e 10 µg/mL do extrato hidroalcoólico das folhas de QG foram administrados por períodos de 24, 48, 72 e 96h. Após cada período, a viabilidade celular foi avaliada pelo método de redução de MTT e a análise da expressão das moléculas foi feita por meio da técnica de imunofluorescência. Os resultados mostram que o extrato de QG não promove redução da viabilidade celular de fibroblastos e pré-osteoblastos em concentrações até 10 µg/mL, nos períodos iniciais (24 e 48h). Porém, uma redução significativa da viabilidade pode ser verificada nos períodos de 72h e 96h para os fibroblastos e 96h para os pré-osteoblastos, expostos a mais alta concentração do extrato (10 µg/mL). O ensaio de imunofluorescência indica que o extrato, nas concentrações de 0.1; 1.0 e 10 µg/mL foi capaz de aumentar a expressão de MMP-14 e HIF-1alpha, em ambos os tipos celulares. Em conclusão, nossos resultados indicam que o extrato de QG exerce um efeito capaz de aumentar a expressão das duas moléculas em estudo (MMP-14 e HIF-1alpha), tanto para os fibroblastos da linhagem NIH/3T3 como para os pré- osteoblastos da linhagem MC3T3-E1. Assim, os compostos de QG podem apresentar potencial para serem utilizados como agentes terapêuticos moduladores da angiogênese, por meio do aumento da expressão de MMP-14 e HIF-1alpha.(AU)

The vegetable specie Qualea grandiflora (QG), popularly known as "pau-ferro", "pauterra-da-folha-grande", "pau-terra" or "pau-de-tucano", very common in the Brazilian Cerrado, is well known due to its varied therapeutic properties. Its indications include preventive actions in the appearance of lesions of gastric mucosa, analgesic, antibacterial, anti-inflammatory and antifungal effects. Thus, QG components could have some action on molecules widely involved in angiogenic and developmental / repair processes, such as Matrix metalloproteinase 14 (MMP-14) and HypoxiaInducible Factor-1α (HIF-1alpha). Thus, the objective of our study was to investigate the effects of QG hydroalcoholic extract on cell viability and expression of MMP-14 and HIF-1alpha in NIH/3T3 fibroblasts and MC3T3-E1 pre-osteoblasts cell lines. For the cell viability assay and expression of the molecules, concentrations of 0.1; 1.0 and 10 µg / mL of the hydroalcoholic extract of leaves of QG, were administered for periods of 24, 48, 72 and 96h. After each period, the cell viability was evaluated by MTT assay and the expression of the molecules was analyzed using the immunofluorescence technique. The results show that the QG extract does not promote reduction of the cellular viability of fibroblasts and pre-osteoblasts in concentrations up to 10 µg/mL in the initial periods (24 and 48h). However, a significant reduction in viability can be observed in 72h and 96h for fibroblasts and 96h for pre-osteoblasts exposed to the highest extract concentration (10 µg/mL). The immunofluorescence assay indicates that the extract, at concentrations of 0.1; 1.0 and 10 µg/mL was able to increase the expression of MMP-14 and HIF-1alpha in both cell types. In conclusion, our results indicate that the QG extract exerts an effect capable of increasing the expression of the two molecules under study (MMP-14 and HIF-1alpha) both for the NIH/3T3 fibroblasts as well as for the MC3T3-E1 pre-osteoblasts cells. Thus, the QG compounds could have potential to be used as angiogenesis modulating therapeutic agents, by increasing the expression of MMP-14 and HIF-1alpha.(AU)

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Period2 is a core circadian gene, which not only maintains the circadian rhythm of cells but also regulates some organic functions. We investigated the effects of mPeriod2 (mPer2) expression on radiosensitivity in normal mouse cells exposed to 60Co-gamma-rays. NIH 3T3 cells were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce endogenous mPer2 expression or transfected with pcDNA3.1(+)-mPer2 and irradiated with 60Co-gamma-rays, and then analyzed by several methods such as flow cytometry, colony formation assay, RT-PCR, and immunohistochemistry. Flow cytometry and colony formation assay revealed that irradiated NIH 3T3 cells expressing high levels of mPer2 showed a lower death rate (TPA: 24 h 4.3% vs 12 h 6.8% and control 9.4%; transfection: pcDNA3.1-mPer2 3.7% vs pcDNA3.1 11.3% and control 8.2%), more proliferation and clonogenic survival (TPA: 121.7 +/- 6.51 vs 66.0 +/- 3.51 and 67.7 +/- 7.37; transfection: 121.7 +/- 6.50 vs 65.3 +/- 3.51 and 69.0 +/- 4.58) both when treated with TPA and transfected with mPer2. RT-PCR analysis showed an increased expression of bax, bcl-2, p53, c-myc, mre11, and nbs1, and an increased proportionality of bcl-2/bax in the irradiated cells at peak mPer2 expression compared with cells at trough mPer2 expression and control cells. However, no significant difference in rad50 expression was observed among the three groups of cells. Immunohistochemistry also showed increased protein levels of P53, BAX and proliferating cell nuclear antigen in irradiated cells with peak mPer2 levels. Thus, high expression of the circadian gene mPer2 may reduce the radiosensitivity of NIH 3T3 cells. For this effect, mPer2 may directly or indirectly regulate the expressions of cell proliferation- and apoptosis-related genes and DNA repair-related genes.

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Cyanoacrylate has been used in medicine and dentistry for many years. It has been used as a postextraction dressing and retrograde filling material in endodontic surgery. The aim of this study was to evaluate the cytotoxic effects of Histoacryl and other two homologue ethyl cyanoacrylates, Super Bonder and Ultrabond, on cultured fibroblasts, using the Trypan blue dye exclusion assay. The cyanoacrylates were applied to round glass coverslips, which were placed in contact with NIH 3T3 cells. After 0, 6, 12 and 24 h (short-term assay; viability) and 1, 3, 5 and 7 days (long-term assay; survival), the cells were examined under phase light microscopy and counted. The data were compared by the Kruskal-Wallis test. In the short-term experiments, only the cultures of the Ultrabond group (GIV) presented significant smaller percentages of cell viability than the cultures of the other groups (GI: control; GII: Super Bonder; GIII: Histoacryl). Although the cultures of the Super Bonder group (GII) presented smaller percentages of cell viability than cultures of the other groups (GI, GIII, GIV) at the long-term assay, this group was the only experimental group presenting a continuous and progressive cell growth. Our results have shown an in vitro biocompatibility of Histoacryl and ethyl cyanoacrylate homologues. These cyanoacrylates could therefore be of importance for endodontic purposes.

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The cytotoxicity of two nitroheterocyclic compounds (NHCD), Nitracrine, 1-nitro-9(3'3'-dimethylaminopropylamino) acridine (Polfa, Poland) and Quinifuryl, 2-(5'-nitro-2'-furanyl) ethenyl-4-[N-[4-(N,N-diethylamino)-1'-methylbutyl] carbamoyl] quinoline (Dr. N. M. Sukhova, Institute of Organic Synthesis, Riga, Latvian Republic), towards two lines of leukaemic cells and a line of non-transformed cells, was determined under normoxia conditions. Although both drugs showed significant cytotoxicity to all cell lines (LC(50) for 24h, < or = 2 microM) with that of Nitracrine exceeding Quinifuryl, their toxicity towards murine leukaemia P388 was substantially higher, compared to murine fibroblasts NIH3T3. In addition, the rate of cell death was also two- to three-fold higher in case of P388 cells versus NIH3T3. Interestingly, human erythroleukaemia K562 cells were shown to uptake the drugs 10 min after their addition to the tissue culture medium, while the LC(50) values were reached after a substantial delay of 3h. This delay might be due to the intracellular transformation of drugs required for cell killing.

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