Zofenopril antitumor activity in mice bearing Ehrlich solid carcinoma: Modulation of PI3K/AKT signaling pathway

Angiotensin-II (AgII) is thought to be crucial for tumor growth and progression. Moreover, hydrogen sulfide (H2S) performs a controversial action in cancer pathology. Zofenopril (ZF) is an angiotensin-converting enzyme (ACE) inhibitor with H2S donating properties. Hence, this study aims at investigating the tumor suppressor activity of ZF and elucidating the involved trajectories in Ehrlich's solid tumor (EST)-bearing mice. EST was induced by the intradermal injection of Ehrlich's ascites carcinoma cells into femoral region. All parameters were assessed after 28 days post-inoculation or one-week thereafter. ZF treatment resulted in significant reduction of tumor weights with marked decrease in IL-6 and VEGF levels in serum, and tumor Ag II and CEA contents. Additionally, the administration of ZF downregulated the tumor gene expression of cyclin-D, ACE-1, and Bcl2 and upregulated the proapoptotic gene, BAX. Moreover, ZF increased CBS gene expression, which is a major contributor to cellular H2S production. In addition, ZF was able to reduce the protein expression of PI3K, pAKT, pGSK-3ß, and NFκB. Our study has provided novel insights into the possible mechanisms by which ZF may produce its tumor defeating properties. These intersecting trajectories involve the interference between PI3K/Akt and CBS signaling pathways

Investigation of effect of paclitaxel on netrin 1 and factor 8 in Ehrlich solid tumors / Investigación del efecto del paclitaxel en netrina 1 y factor 8 en tumores sólidos de Ehrlich

SUMMARY: Cancer known as a malignant tumor, is a class of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. The Ehrlich tumor is a mammary adenocarcinoma of mice developed in solid and ascitic forms. This study was aimed to investigate the effects of paclitaxel on Netrin 1 and Factor 8 expression and also in tumor cell proliferation, apoptosis, angiogenesis, and development of tumor in Ehrlich solid tumors treated with paclitaxel. In this study, 26 adult Balb/C male mice were used. 6 of them were used as stock. Ehrlich ascites cells taken from animals in stock were injected subcutaneously from the neck area to all animals. The mice were randomly assigned to two groups of ten rats per group. Paclitaxel treatment group 10 mg/kg were administered to mice intraperitoneally (i.p.) 4,9, and 14th days. 15th day the animals were sacrificed and tumor tissues were taken. Paraffin-embedded solid tumor sections were stained Hematoxylin & Eosin, Masson's Trichrome. Also solid tumor sections were stained immunohistochemically with Netrin1 and Factor 8. Tunel method was applied to determine apoptosis. Paclitaxel applied as a therapeutic Ehrlich solid tumor reduced the volume of tumors in the treatment groups. At the end of the experiments, in the treatment groups' significantly reduced the Netrin 1 expression and microvessel density compared to the group control. Also paclitaxel in the treatment group increased the number of apoptotic cells. We suggest that decreasing the expression of Netrin 1 would be reduced vessel density and increased apoptosis.

RESUMEN: El cáncer, conocido como tumor maligno, es una clase de enfermedad que involucra un crecimiento celular anormal con potencial de invadir o diseminarse a otras partes del cuerpo. El tumor de Ehrlich es un adenocarcinoma mamario de ratones desarrollado en formas sólidas y ascíticas. Este estudio tuvo como objetivo investigar los efectos del paclitaxel en la expresión de Netrin 1 y Factor 8 y también en la proliferación de células tumorales, apoptosis, angiogénesis y desarrollo de tumores sólidos de Ehrlich tratados con paclitaxel. En esta investigación se utilizaron 26 ratones machos Balb / C adultos. Seis de ellos se utilizaron como stock. Se inyectaron por vía subcutánea células de ascitis de Ehrlich tomadas de animales en la zona del cuello. Los ratones se asignaron aleatoriamente a dos grupos de diez ratas por grupo. Se administraron 10 mg/kg del grupo de tratamiento con paclitaxel a ratones por vía intraperitoneal (i.p.) 4, 9 y 14 días. El día 15 se sacrificaron los animales y se extrajeron los tejidos tumorales. Las secciones de tumor sólido incluidas en parafina se tiñeron con hematoxilina y eosina y tricrómico de Masson. También se tiñeron inmunohisto-químicamente secciones de tumor sólido con Netrin1 y Factor 8. Se aplicó el método Tunel para determinar la apoptosis. El paclitaxel aplicado como tumor sólido terapéutico de Ehrlich redujo el volumen de tumores en los grupos de tratamiento. Al final de los experimentos, en los grupos de tratamiento se redujo significativamente la expresión de Netrin 1 y la densidad de microvasos en comparación con el grupo control. Además, el paclitaxel en el grupo tratamiento aumentó el número de células apoptóticas. Sugerimos que la disminución de la expresión de Netrin 1 reduciría la densidad de los vasos y aumentaría la apoptosis.

Enhancing Antitumor Efficacy of Cisplatin Low Dose by EDTA in Ehrlich Ascetic Carcinoma Bearing Mice

Abstract In a recent study, the treatment of different human cancer cell lines in vitro with ethylene diamine tetra-acetic acid (EDTA) showed a promising anticancer activity which could be a novel promising approach for cancer treatment. The aim of this study is to address the ability of EDTA to enhance the antitumor efficacy of the low dose of cisplatin (Cis) treatment in Ehrlich ascetic carcinoma (EAC) bearing mice. Sixty female albino mice were divided into six groups. The 1st group of mice was served as a negative control. 2nd - 6th groups were inoculated intraperitoneal (i.p) with 2×106 EAC cells/mouse. After one day of inoculation, the 2nd, 3rd and 4th groups were injected daily for 6 days (early treatment) with phosphate buffer saline, low dose of Cis and Cis/EDTA, respectively. After six days, the 5th and 6th groups were injected with the low dose of Cis and Cis/EDTA for 6 consecutive days (late treatment), respectively. At day 14, all groups of mice were sacrificed, sera were collected for biochemical assessment, then tumor volumes, counts, live and dead cells were determined from all groups. The results showed that EDTA co-treatment enhanced the efficacy of low dose of Cis at early and late time points. In addition, EDTA co-treatment potentially ameliorated the Cis-induced side effects on liver and kidney functions. In summary, co-therapy with EDTA could enhance the chemotherapeutic efficacy of low dose of Cis.

Natural killer activity of the spleen cells of Ehrlich tumor-bearing mice treated with Copaifera multijuga extract / Atividade natural killer das células esplênicas de camundongos portadores do tumor de Ehrlich tratados com extrato de Copaifera multijuga

AIMS: Copaifera multijuga Hayne oleoresin is commonly used in traditional medicine owing to its anti-inflammatory, antiseptic, antitumor, and antibacterial properties. However, little is known about the effect of the compounds from the bark of this plant. In this study, the immunomodulatory effect of the ethanolic extract of C. multijuga bark via natural killer activity of non-adherent spleen cells of Ehrlich tumor-bearing mice was evaluated. METHODS: Male Swiss mice were inoculated subcutaneously with 1×106 Ehrlich tumor cells (Ehrlich and Ehrlich/C. multijuga group) or phosphate buffered saline solution (control group and C. multijuga group) and treated orally daily with C. multijuga extract (200 mg kg-1, 0.1 mL per mouse, for the Ehrlich/C. multijuga and C. multijuga groups) or phosphate buffered saline solution (control group and Ehrlich group). The four experimental groups consisted in eight mice each and were organized in two sets, one treated for seven days and another treated for 14 days, totalizing 64 mice throughout the experiment. Twenty-four hours after the last oral administration, the mice were euthanized and the spleen tissue was isolated to prepare a non-adherent spleen cell suspension and to evaluate natural killer activity. Data are presented as the cell lysis percentage of Yac.1 target cells by non-adherent spleen cells. RESULTS: Treatment for seven days increased natural killer activity in the Ehrlich/C. multijuga group (21.20±8.89, p<0.05) compared to the control group (3.14±2.71, p<0.05); however, this effect was not maintained in the groups treated for 14 days (Control: 6.02±6.98, Ehrlich: 4.82±7.72, C. multijuga: 2.07±2.10, Ehrlich/C. multijuga: 2.01±1.63, p>0.05). CONCLUSIONS: Treatment for seven days with an ethanolic extract of C. multijuga bark enhanced the natural killer activity of non-adherent spleen cells from Ehrlich tumor-bearing mice.

OBJETIVOS: O óleo-resina de Copaifera multijuga Hayne é popularmente utilizado na medicina tradicional por suas propriedades antiinflamatória, antisséptica, antitumoral e antibacteriana. Entretanto, há poucos estudos sobre o efeito dos compostos obtidos da casca da planta. Assim, o objetivo deste trabalho foi avaliar o efeito imunomodulador do extrato etanólico da casca da C. multijuga na atividade natural killer das células esplênicas não aderentes de animais portadores do tumor de Ehrlich. MÉTODOS: Camundongos Swiss machos foram inoculados subcutaneamente com 1×106 células do tumor de Ehrlich (grupo Ehrlich e Ehrlich/C. multijuga) ou com solução salina tamponada (grupo controle e C. multijuga) e tratados diariamente (gavagem) com extrato de C. multijuga (200 mg kg-1, 0,1mL cada um, para os grupos Ehrlich/C. multijuga e C. multijuga) ou com solução salina tamponada (grupo controle e grupo Ehrlich). Os quatro grupos experimentais consistiram de oito animais cada e foram organizados em dois conjuntos, um tratado por sete dias e outro tratado por 14 dias, totalizando 64 animais durante todo o experimento. Após 24 horas do término do tratamento, os animais foram eutanasiados para obtenção da suspensão de células esplênicas não aderentes e avaliação da atividade natural killer. Os resultados são apresentados como porcentagem da lise celular das células alvo Yac.1 pelas células esplênicas não aderentes. RESULTADOS: A análise dos resultados demonstrou que aos sete dias de tratamento a C. multijuga aumentou a atividade natural killer no grupo Ehrlich/C. multijuga (21,20±8,89, p<0,05) em comparação ao grupo controle (3,14±2,71, p<0,05), entretanto tal efeito não foi mantido nos grupos tratados por 14 dias (Controle: 6,02±6,98; EHR: 4,82±7,72; C. multijuga: 2,07±2,10; EHR/C. multijuga: 2,01±1,63, p>0,05). CONCLUSÕES: Os resultados demonstraram que o tratamento com o extrato etanólico da casca de C. multijuga favoreceu a atividade natural killer das células esplênicas não aderentes dos animais portadores do tumor de Ehrlich apenas aos sete dias de tratamento.

Walker-256 Tumor: Experimental Model, Implantation Sites and Number of Cells for Ascitic and Solid Tumor Development

Abstract The Walker-256 tumor is an important experimental model that allow the development of therapies as the biological behavior of this tumor is similar that occur in humans. In front of the above considerations, the aim of this study was to describe the experimental model of Walker-256 tumor, identify the implantations sites as well as define a usual quantity of tumoral cells to induce the ascitic and solid tumor, according to the specialized literature. Were selected 45 articles using the keyword "Walker-256 tumor", free available. Were possible to observe that 58% (n=26) of the studies inoculate the tumor cells in the animals flank 33% (n = 15) in the tibia bone, 7% (n = 3) in the femur and 2% (n = 1) in the paw. The major quantitates of cells used were 8 x 107 (20%), 1 x 105 (13%), 1 x 106 (11%) and 2 x 107 (11%). After that, the site commonly used to inoculate was the flank and quantitate still a controversy, being 1x105 and 8x107 the concentrations more used.

Atividade antitumoral da melatonina sobre o tumor de ehrlich implantado em camundongos swiss / Melatonin antitumor activity on Ehrlich tumor implanted in Swiss mice

Introdução: Um mecanismo proposto para a ação oncostática da melatonina no tumor de mama é a inibição do hormônio liberador das gonadotrofinas (GnRH), dificultando assim a liberação dos hormônios luteinizante (LH) e folículo estimulante (FSH) e, consequentemente, reduzindo a produção ovariana de estradiol. Objetivo: Avaliar a atividade antitumoral da melatonina sobre o tumor de Ehrlich em camundongos fêmeas Swiss, através de sua interação com o aparelho reprodutor. Método: Foram utilizados 56 camundongos fêmeas Swiss (Mus musculus), de 60 dias de idade, divididas em sete grupos experimentais (n = 8 animais/ grupo): A- Pinealectomizados tratados com melatonina; B- Shampinealectomizados tratados com melatonina; C- Pinealectomizados sem tratamento; D- Sham-pinealectomizados sem tratamento; EPinealectomizados tratados com veículo da melatonina; F- Shampinealectomizados tratados com veículo da melatonina; G- Controle. Resultados e Discussão: Alterações ovarianas foram significativas entre os grupos, observando-se um aumento no peso dos ovários e no número de folículos ovarianos dos animais pinealectomizados, enquanto os animais sham-pinealectomizados apresentaram um maior número de corpos lúteos. Os animais pinealectomizados apresentaram maior taxa de crescimento tumoral quando comparados aos animais tratados com melatonina, tendo estes últimos apresentado também uma percentagem de inibição tumoral. Conclusão: A análise destes resultados nos permite concluir que a melatonina, através de sua atuação no ovário, interfere nos mecanismos regulatórios dos processos de crescimento do tumor de Ehrlich. Pode-se conjecturar que eventuais alterações nos padrões de ritmicidade endógena para a melatonina pode predispor os organismos a uma maior incidência de tumor.

Introduction: A proposed mechanism for the oncostatic action of melatonin in breast tumors is the inhibition of gonadotropinreleasing hormone (GnRH), thus hindering the release of luteinizing hormones (LH) and stimulating follicle (FSH) and, consequently, reducing ovarian estradiol production. Objective: This study aim was to evaluate the melatonin antitumor activity on the Ehrlich tumor in female Swiss mice, through its interaction with the reproductive system. Method: we used 56 female Swiss mice (Mus musculus), 60 days of age, were divided into seven experimental groups (n = 8 animals/group): A- Pinealectomized melatonin treated; B- Shampinealectomized melatonin treated; C- Pinealectomized no treated; D- Sham-Pinealectomized no treated; E- Pinealectomized melatonin vehicle treated; F- Sham-pinealectomized melatonin vehicle treated; G- Control. Results and discussion: Ovarian changes were significant between the groups, observing an increase in ovarian weight and follicles number in pinealectomized animals, whereas the sham-pinealectomized animals exhibited greater numbers of corpus luteum. The pinealectomized animals had higher rate of tumor growth than melatonin treated animals, the last having also presented a percent inhibition of the tumor. Conclusion: Analysis of these results allows us to conclude that melatonin, through its actions in the ovary interfere with regulatory mechanisms of the processes of Ehrlich tumor growth. May conjecture that any changes in the patterns of endogenous rhythmicity for melatonin, can predispose organisms to a higher incidence of tumor.

High-diluted thymulin on Ehrlich tumor growth in mice and the importance of tumor microenvironment

Introduction: The aim of the present study was to describe different biological aspects of Ehrlich tumor in mice, such as body weight evolution, tumor growth rate, histological organization and systemic immune response after treatment with high-diluted thymulin (10-9 M, named 5CH). Methods: Tumor assessment was focused on macro- and microscopic aspects; parameters included occurrence of necrosis, embolism and tumor development, in addition to quantitative analysis of apoptosis (caspase-3), cell proliferation (Ki-67) and angiogenesis (vascular endothelial growth factor - VEGF) by means of specific immunohistochemistry markers. Spleen cell populations were evaluated by flow cytometry analysis. Results: Mice treated with thymulin 5CH exhibited changes in the tumor microenvironment, such as reduced micro-embolism incidence and cytokeratin expression, with increased caspase-3 expression in the tumor cells. These findings indicate some apoptotic activity by the tumor cells induced by the treatment, even though no reduction of the macroscopic tumor mass occurred. No changes in the systemic immune response were detected, as the balance among spleen cell populations remained unchanged. Conclusions: The results indicate that treatment of mice bearing Ehrlich tumor with thymulin 5CH induces some specific changes in the tumor environment. However, it did not influence systemic immunity parameters. Adjuvant use of thymulin 5CH in oncological clinical practice is still a matter of discussion. (AU)

Assessment of electrochemotherapy effects on the development of Ehrlich solid tumor in swiss mice using a novel electroporator device / Avaliação dos efeitos da eletroquimioterapia sobre o desenvolvimento do tumor sólido de Ehrlich em camundongos Swiss utilizando um novo eletroporador

Electrochemotherapy is a local anticancer treatment in which non-permeant chemotherapeutic drugs are associated with electric pulses of well-established parameters. The electric pulses cause pores to open on the plasma membrane and facilitate drug transport, enhancing cytotoxicity and reducing side effects. Assessment of electrochemotherapy effects on Ehrlich solid tumor development in this work aims to evaluate in vivo usage of the electroporator device developed by the Department of Electrical Engineering of Engineering School of UFMG. Therefore, 40 Swiss mice were inoculated with Ehrlich tumor cells, and developed the tumor in solid form. After 21 days, mice were subjected to specific treatment protocols (control, bleomycin, electric pulses and electrochemotherapy); 17 days later they were euthanized and the tumors collected for histopathology analysis. Electrochemotherapy induced discrete weight loss and an inflammatory response in the tumor, which was not seen on the other treatment groups. Bleomycin alone induced necrosis. Both groups showed lower cellular proliferation rates. From this study, it was concluded that the animals tolerated electrochemotherapy treatment under anesthesia and the electroporator device developed by the Engineering School of UFMG was adequate when used in an electrochemotherapy protocol.(AU)

Eletroquimioterapia é uma modalidade de tratamento local contra o câncer em que a administração de quimioterápicos não penetrantes à membrana plasmática é associada à aplicação de pulsos elétricos com parâmetros bem estabelecidos, que abrem poros na membrana plasmática e facilitam a entrada desses fármacos nas células, aumentando sua citotoxicidade e reduzindo efeitos colaterais. A avaliação dos efeitos da eletroquimioterapia sobre o desenvolvimento do tumor sólido de Ehrlich em camundongos Swiss neste trabalho teve como objetivo testar o uso in vivo do aparelho eletroporador desenvolvido pelo Departamento de Engenharia Elétrica da Escola de Engenharia da UFMG. Para tanto, foram utilizados 40 camundongos fêmeas da linhagem Swiss, nos quais foram inoculadas células de tumor de Ehrlich, para o desenvolvimento do tumor na forma sólida. Após 21 dias, os camundongos foram submetidos ao protocolo de tratamento específico (controle, bleomicina, pulsos elétricos e eletroquimioterapia); 17 dias depois foram eutanasiados e seus tumores coletados para análise histopatológica e imuno-histoquímica. A eletroquimioterapia induziu perda de peso discreta e uma resposta inflamatória no tumor que não foi observada nos outros grupos. O grupo bleomicina apresentou maior porcentagem de necrose. Ambos os grupos apresentaram menor índice de proliferação celular. Com este estudo, pode-se concluir que o tratamento sob anestesia foi bem tolerado pelos animais e que o aparelho eletroporador desenvolvido pela Escola de Engenharia da UFMG é adequado para utilização em um protocolo de eletroquimioterapia.(AU)

Synthesis and evaluation of platinum complexes with potential antitumor activity

Abstract A novel series of platinum (II) complexes was synthesized and the complexes were evaluated for their in vitro cytotoxicity against four human cancer cells lines. Five platinum complexes showed activity against at least one tumor cell line. Complexes 3 and 6 were promising, being active, at micromolar concentrations, against all the assayed tumor cell lines. Compound 3 was selected for further studies in mice with Ehrlich solid tumors and it was able to reduce the rate of tumor growth significantly during the first seven days. However, at the end of the experiments, there was no significant difference between the group of animals treated with 3 and the control group. The low solubility of the compound in the assay conditions can explain, at least in part, these results.

Growth factors and cox2 in wound healing: an experimental study with ehrlich tumors / Avaliação dos fatores de crescimento e cox-2 na cicatrização de camundongos com neoplasia de ehrlich

ABSTRACT Background: Healing is an innate biological phenomenon, and carcinogenesis acquired, but with common humoral and cellular elements. Carcinogenesis interferes negatively in healing. Aim: To evaluate the histological changes in laparotomy scars of healthy Balb/c mice and with an Ehrlich tumor in its various forms of presentation. Methods: Fifty-four mice were divided into three groups of 18 animals. First group was the control; the second had Ehrlich tumor with ascites; and the third had the subcutaneous form of this tumor. Seven days after tumor inoculation, all 54 mice were submitted to laparotomy. All of the animals in the experiment were operated on again on 7th day after surgery, with resection of the scar and subsequent euthanasia of the animal. The scars were sent for histological assessment using immunohistochemical techniques to evaluate Cox-2 (cyclooxygenase 2), VEGF (vascular endothelial growth factor) and FGF (fibroblast growth factor). Semi-quantitatively analysis was done in the laparotomy scars and in the abdominal walls far away from the site of the operation. Results: Assessing the weight of the animals, the correct inoculation of the tumor and weight gain in the group with tumoral ascites was observed. The histological studies showed that groups with the tumor showed a statistically significant higher presence of Cox-2 compared to the control. In the Cox-2 analysis of the abdominal wall, the ascites group showed the most significant difference. VEGF did not present any significant differences between the three groups, regardless of the site. The FGF showed a significant increase in animals with the tumor. Conclusion: Histological findings in both laparotomy scar and the abdominal wall showed that with Ehrlich's neoplasia there was an exacerbated inflammatory response, translated by more intense expression of Cox-2 and greater fibroblast proliferation, translated by more intense expression of FGF, that is, it stimulated both the immediate inflammatory reactions, observed with Cox-2 reactions, and late scarring by fibroblasts and FGF.

RESUMO Racional: A cicatrização é fenômeno biológico inato, e a carcinogênese adquirido, mas com elementos humorais e celulares comuns. A carcinogênese interfere de forma negativa na cicatrização. Objetivo: Avaliar as modificações histológicas nas cicatrizes laparotômicas de camundongos Balb/c sadios como controles, e com a neoplasia de Ehrlich, em suas diferentes formas de apresentação. Métodos: Foram utilizados 54 camundongos, divididos em três grupos de 18 animais cada um. O primeiro era controle; o segundo com a neoplasia de Ehrlich em sua forma ascítica; e o terceiro na forma subcutânea. Sete dias após a inoculação do tumor, todos os 54 camundongos foram submetidos à laparotomia e reoperados no sétimo dia de pós-operatório, com ressecção da cicatriz e posterior eutanásia. As cicatrizes foram encaminhadas para estudo histológico com técnicas imunoistoquímicas para avaliar Cox-2 (ciclo-oxigenase 2), VEGF (fator de crescimento do endotélio vascular) e FGF (fator de crescimento dos fibroblastos) e analisadas de forma semiquantitativana tanto na cicatriz laparotômica como na parede abdominal mais distante do local operado. Resultados: Avaliando o peso, observou-se a correta inoculação do tumor e o aumento de peso no grupo com a neoplasia na modalidade ascítica. Os estudos histológicos mostraram que os grupos com a neoplasia apresentaram maior presença da Cox-2 em relação ao controle, estatisticamente significante. No estudo da Cox-2 da parede abdominal foi o local em que o grupo ascítico apresentou a diferença mais expressiva. O VEGF não apresentou diferenças significantes entre os três grupos, independentemente do local estudado. O FGF teve aumento significante nos animais com neoplasia. Conclusão: Os achados histológicos encontrados tanto na cicatriz das laparotomias quanto na parede abdominal mostraram que com a neoplasia de Ehrlich houve resposta inflamatória exacerbada, traduzida por expressão mais intensa da Cox-2 e maior proliferação fibroblástica, traduzida por expressão mais intensa do FGF, ou seja, estimulou tanto as reações inflamatórias imediatas, observadas nas reações da Cox-2, como nas cicatriciais tardias com os fibroblastos e o FGF.

Evaluation of pro-apoptotic activity of Chelidonium majus L. and/or gamma-radiation against ehrlich as carcinoma transplanted in mice / Avaliação da atividade pró-apoptótica da Chelidonium majus L. e/ou radiação gama contra o tumor ascítico de ehrlich transplantado em ratos

This study was to evaluate, the pro-apoptotic activity of ethanolic Chelidonium majus L. (CM) leaf extract and/or gamma-radiation in Ehrlich ascites carcinoma (EAC) bearing mice by measuring tumor volume, apoptotic factors (caspase-3, Bcl-2 and Bax proteins, tumor necrosis factor-alpha (TNF-), angiogenesis factors (matrix metalloproteinase (MMP-2 and MMP-9); tissue inhibitor of metalloproteinases (TIMP-1), DNA fragmentation, besides histopathological examination. After tumor inoculation, mice received CM (100 mg/body weight/day) on the 10th day and were exposed to whole body -radiation (6Gy) on the 17th day. The data obtained reveales that combining CM with - radiation exposure induce significant regression in tumor growth, up-regulate Bax, caspase-3, TIMP-1 and inhibit Bcl-2, TNF-, MMP-(2 and 9). Cytotoxicity is substantiated by increased DNA-fragmentation (comet assay and DNA content) and histological examination. It could be suggested that combining CM with gamma-radiation increased apoptosis of tumor cells which would help to increase the lifespan of mice.

Este estudo foi realizado para avaliar a atividade pró-apoptótica do extrato etanólico da folha da Chelidonium majus L. (CM) e/ou radiação gama no tumor ascítico de Ehrlich tendo ratos para a medição do volume do tumor, fatores apoptóticos (caspase-3, proteínas Bcl-2 e Bax, fator de necrose tumoral (TNF-)), fatores angiogênicos (metaloproteinase matriz (MMP-2 e MMP-9)); inibidor tecidual de metaloproteinase (TIMP-1), fragmentação de DNA, além de exame histopatológico. Depois da inoculação do tumor, os ratos receberam CM 100 mg/peso corporal/dia no 10º dia e tiverem o corpo completo exposto a radiação gama (6Gy) no 17º dia. Os dados obtidos revelam que combinar CM com exposição à radiação gama induz a uma regressão significativa no crescimento do tumor, regula positivamente Bax, caspase-3, TIMP-1 e inibe Bcl-2, TNF-, MMP-(2 e 9). Citotoxicidade é substanciada pelo aumento da fragmentação do DNA (ensaio cometa e conteúdo de DNA) e exame histopatológico. Poderia ser sugerido que combinar CM com radiação gama aumentou a apoptose das células tumorais o que ajudaria a aumentar a expectativa de vida dos ratos.

Use of raw Euphorbia tirucalli extract for inhibition of ascitic Ehrlich tumor / Avaliação do uso do extrato bruto de Euphorbia tirucalli na inibição do tumor ascítico de ehrlich

Objective: to evaluate the effect of the Euphorbia tirucalli hydroalcoholic extract (ETHE) on the development of Ehrlich Tumor, in its ascitic form. Methods: we intraperitoneally inoculated 15 Swiss mice with 10.44 x 107 cells of Ehrlich Tumor and divided them in two groups one day after: ETHE Group (eight mice), treated with a dosage of 125 mg/kg/day of EHTE for five days; and Control Group (seven mice), treated only with 0.9% isotonic saline solution over the same period. The treatment was done by gavage. Ten days after inoculation, four mice from each group were sacrificed for quantification of tumor cell number, ascitic fluid volume and bone marrow cell number. The remaining animals were maintained to evaluate survival. Results: The ascitic fluid volume and the tumor cell number were decreased in the ETHE group when compared with the control group, but with no statistical significance. On the other hand, survival was higher in the ETHE group, as well as the number of bone marrow cells. Conclusion: Treatment with ETHE after inoculation of Ehrlich Tumor decreases its development and increases survival and the bone marrow cellularity, thus reducing the myelosuppression present in the Ehrlich Tumor bearing mice.

Objetivo: avaliar o efeito do extrato hidroalcoólico de Euphorbia tirucalli (ETHE) sobre o desenvolvimento do tumor de Ehrlich em sua forma ascítica. Métodos: quinze camundongos Swiss foram inoculados via intraperitoneal com 10,44x107 células do tumor de Ehrlich e um dia depois foram divididos em dois grupos: Grupo ETHE (oito camundongos), tratados com a dose de 125mg/kg/dia de ETHE por cinco dias e Grupo Controle (sete camundongos), tratado apenas com 0,9% de solução salina isotônica em relação ao mesmo período. O tratamento foi realizado por gavagem. Dez dias após a inoculação, quatro animais de cada grupo foram sacrificados para a quantificação do número de células de tumor, do volume de fluido ascítico e do número de células da medula óssea. Os demais animais foram mantidos, para avaliar a sobrevivência. Resultados : o volume de líquido ascítico e do número de células tumorais foram menores no grupo ETHE quando comparado ao grupo controle, porém sem significância estatística. Por outro lado, a sobrevivência dos animais foi maior no grupo de ETHE, bem como, a quantidade de células de medula óssea. Conclusão: o tratamento com ETHE, após a inoculação do tumor, diminuiu o seu desenvolvimento e aumentou sobrevida, bem como, a celularidade da medula óssea, reduzindo assim, a mielossupressão presente nos animais portadores de tumor de Ehrlich.

Effects of different extracts of the mushroom Agaricus blazei Murill on the hematologic profile of mice with Ehrlich tumor / Efeitos de diferentes extratos do cogumelo Agaricus blazei Murill sobre o perfil hematológico de camundongos com tumor de Ehrlich

The aim of this study was to investigate the effect of the mushroom Agaricus blazeii Murril (ABM) extracts on the hematological profile of Swiss mice bearing an Ehrlich solid tumor. Three fractions (total extract, polysaccharides, and supernatant) of ABM extracts obtained by four methods (ultrasonic or water bath, at pH 4 or pH 7) were administered to mice over 21 days. Polysaccharide solutions were analyzed by gas and liquid chromatography that showed both mannose and glucose concentrations. The method of extraction influenced the degree of glucose polymerization and the mannose/glucose relationship. The treatment with ABM supernatant at pH 7 and water bath was associated with reduced concentrations of leukocytes and lymphocytes and altered the percentage of CD4+ and CD8+ lymphocytes in Ehrlich tumor-bearing mice. The treatment with the ABM extract in water bath and ultrasound at pH 4 resulted in lower lymphocyte counts, regardless of tumor presence, and greater granulocyte values in mice with Ehrlich tumor than in controls. We concluded that different fractions and methods of extraction of A. blazei produced differing blood profiles in mice inoculated with the Ehrlich tumor.

O objetivo deste estudo foi investigar o efeito de diferentes extratos do cogumelo Agaricus blazeii Murril (ABM) sobre o perfil hematológico de camundongos Swiss portadores de tumor de Ehrlich sólido. Três frações (extrato total, polissacarídeos e sobrenadante) dos extratos de ABM foram obtidas por quatro métodos (sonificador, banho-maria, em pH 4 ou pH 7) e administradas para camundongos durante 21 dias. Soluções de polissacarídeos foram analisadas por cromatografia gasosa e líquida, que mostraram concentrações de glucose e manose. O método de extração influenciou o grau de polimerização da glicose e a relação manose/glucose. O tratamento com o sobrenadante de ABM (em pH 7 e banho-maria) estava associado com reduzidas concentrações de leucócitos e linfócitos, além de alterar a porcentagem de linfócitos CD4+ e CD8+ em camundongos portadores de tumor sólido de Ehrlich. O tratamento com extratos de ABM, obtidos tanto em banho-maria como no sonificador em pH 4, resultou nas mais baixas contagens de linfócitos, independentemente da presença do tumor, e nos maiores valores de granulócitos em camundongos com tumor de Ehrlich. Conclui-se que os diferentes métodos de extração com as respectivas frações de A. blazei são capazes de intereferir no perfil hematológico de camundongos com tumor sólido de Ehrlich.

Efeito do tratamento com melatonina sobre a produção de metástases neoplasias / Result of the treatment with melatonin inthe production of neoplasic metastasis

Introdução: Neoplasias são células com proliferação e diferenciação anormais. Quando essas células tornam-se capazes de originar outras células em tecidos e vasos diferentes do inicial são denominadas metástases. A melatonina (N-acetil-5-metoxitripamina) é um hormônio secretado pela glândula pineal e que tem associação com o controle celular tumoral. Objetivo: pretendeu-se avaliar o efeito do tratamento com melatonina sobre a produção e crescimento de metástase linfática do tumor sólido de Ehrlich (TE) em camundongos. Método: foram empregados 14 camundongos distribuídos em 2 grupos: controle (GC, N = 07 / 0,1ml de solução fisiológica, via oral, 1x/dia) e teste (GT, N = 07 / 10mg/kg de melatonina, via oral, 1x/ dia). Todos os animais foram inoculados com 10 6 células tumorais no coxim plantar da pata traseira direita. Resultados e Discussão: D=decorridos 17 dias de tratamento os animais foram eutanasiados, removemos as 02 patas traseiras para avaliar o peso da massa tumoral sendo que o grupo controle apresentou peso maior (GC = 0,62 ± 0,14). Também foi removido o linfonodo poplíteo para mensuração da área, constatando redução significativa no grupo teste (GT = 4,37 ± 1,58). A redução da área do linfonodo pode ser associada a uma redução no número de células tumorais presentes nos linfonodos (GT = 62,8 ± 13,07). Conclusão: Concluímos que o tratamento com melatonina reduziu significativamente o crescimento tumoral e metastático do TE.

Introduction: Neoplasms are cells with unusual proliferation and differentiation. When these cells become able to generate other cells in material and veins different from the beginning one arenamed metastasis. The melatonin (N-acetil-5-methoxytryptamine) is a hormone secreted by the pineal gland and it has association with the tumor cell control. Objective: the general aim was to evaluate the treatment result with melatonin on the metastasis lymphatic production and growth of the Ehrlich Tumor in mice. For this experience, 14 mice were used and shared in 2 groups: control (GC, N = 07 / 0,1ml of sterilizing solution, oral, once a day) and test (GT, N = 07 / 10mg/kg of melatonin, oral, once a day). All the animals were inoculated with 106 tumor cells in the footpad of the right back paw. Results and discussion: after 17 days of treatment the animals were euthanized and 2 back paws were removed to evaluate the tumor bulk weight considering that the control group was heavier (GC = 0,62 ± 0,14). The popliteallymph node was also removed for the area measurement where asmaller area for the test group was observed (GT = 62,8 ± 13,07). Conclusion: we concluded that the treatment with melatonin meaningfully reduced the metastatic growth.

Protective and curative effect of thymoquinone on ehrlich solid carcinoma inoculated mice

Thymoquinone [TQ], the main constituents of the volatile oil from Nigella sativa seeds and it is reported to protect laboratory animals against chemical toxicity and induction of carcinogenesis. This study was undertaken to investigate the potential protective and curative effect of TQ on Ehrlich solid carcinoma cells [ESC] inoculated mice-induced carcinogenesis. [50] Swiss albino mice were divided into five groups. Control group, Thymoquonone group: animals of this group were orally treated with TQ [10 mg/kg/day] for 4 weeks, Ehrlich Carcinoma group: animals of this group were inoculated intramuscularly with 0.2mL Ehrlich Ascites Carcinoma [2.5×10[6] cells] in the right thigh of the lower limb, Thymoquinone and Ehrlich carcinoma group: animals of this group were pre-treated with TQ for 14 days then inoculated with [EAC] and Ehrlich Carcinoma and Thymoquinone group: in this group animals were inoculated with EAC then after 8 days received TQ orally for a month. Morphological, Molecular as well as histopathological and ultrastructural changes were examined. Our results revealed that TQ showed a significant anti-tumor activity in ESC bearing mice represented by a reduction in tumor weight and volume. Flow cytometric analysis illustrated that the level of apoptosis is significantly decreased in ESC inoculated group. Otherwise, TQ+ESC and ESC+TQ groups showed a highly significant increase in apoptosis G0/1 peak. The level of P53 protein expression showed a significant decrease in ESC inoculated group, and this decrease was ameliorated in TQ+ESC and ESC+TQ groups when compared to ESC inoculated group. Histopathological observations showed a reduction in tumor size after treatment with Thymoquinone and this tumor was found to be discontinuous and fragmented with slowly growing. Our results revealed that TQ has potential benefits in the prevention of the onset and progression of solid tumor model in mice

Histopathological evaluation of tumor necrosis and volume after cyanogenic chemotherapy

PURPOSE: To determine the percentage of tumoral necrosis and volume after cyanogenic chemotherapy. METHODS: Histopathological findings of 20 Swiss mice inoculated subcutaneously in the left abdominal wall with 0.05 ml of cell suspension containing 2.5 x 105 viable cells of the Ehrlich tumor were evaluated. The tumor response to cyanogenic chemotherapy was determined using a system that comprises two inhibition factors of tumor growth by calculating the percentage of necrosis in the tumor tissue and calculation of tumor volume in treated animals relative to that in control animals. The importance of this system has been validated by the correlation between tumor inhibition in the groups treated with the respective percentages of necrosis. RESULTS: While the control group presented an average of 13.48 ± 14.71% necrosis and average tumor volume of 16.18 ± 10.94, the treated group had an average of 42.02 ± 11.58 and 6.8 ± 3.57, respectively. The tumor inhibition was significantly associated with treatment (p=0.0189). The analysis of necrosis percentage showed a significant prognostic importance (p=0.0001). CONCLUSION: It is concluded that the effect of cyanogenic chemotherapy showed strong inhibitory action of tumor growth, as well as an increase in its area of necrosis. .

Anti-proliferative and angio-suppressive effect of Stoechospermum marginatum (C. Agardh) Kutzing extract using various experimental models

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

Anti-proliferative and angio-suppressive effect of Stoechospermum marginatum (C. Agardh) Kutzing extract using various experimental models

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

Antioxidant and Antitumor Activity of Plumeria acuminata in Ehrlich Ascites Carcinoma Bearing Swiss Albino Mice.

Aim: This study was designed to determine the antitumor and antioxidant properties of crude methanol extract from the leaves of Plumeria acuminata (Apocynaceae) (MEPA) against Ehrlich Ascites Carcinoma (EAC) bearing Swiss albino mice. Study Design: Study design is methodology, mentioned below. Place and Duration of Study: Division of Pharmacology, Department of Pharmaceutical Technology, Jadavpur University, Jadavpur, Kolkata, India between 2006 and 2007. Methodology: The extract was administered at the doses of 100, 250 and 500 mg/kg per day for 14 days, after 24 hr of tumor inoculation. After the administration of the last dose followed by 18 hr fasting, mice were then sacrificed for observation of antitumor activity. The effect of MEPA on the growth of transplantable murine tumor, life span of EAC bearing host, viable and non-viable cell count, packed cell volume, hematological profile and biochemical parameters such as lipid peroxidation (LPO), reduced glutathione content (GSH), superoxide dismutase (SOD) and catalase (CAT) activities were estimated. Results: MEPA caused significant (P<0.01) decrease in tumor volume, packed cell volume and viable count; and it prolonged the life span of EAC-tumor bearing mice. Hematological studies reveal that the Hb content and RBC count were decreased in EAC treated mice, whereas the restoration to near normal levels was observed in extract treated animals. MEPA significantly (P<0.05) decreased the levels of LPO and significantly increased the levels of GSH, SOD and CAT. Moreover the MEPA was found to be devoid of conspicuous short-term toxicity in the mice when administered daily for 14 days at the doses of 100, 250 and 500 mg/kg Conclusion: The results suggested that the methanol extract of Plumeria acuminata leaves exhibited antitumor effect by modulating lipid peroxidation and augmenting antioxidant defense system in EAC bearing Swiss albino mice.

Sulfane sulfur deficiency in malignant cells, increasing the inhibiting action of acetone cyanohydrin in tumor growth

PURPOSE: To demonstrate the irreversible poisoning action of the acetone cyanohydrin (AC) in malignant cells. METHODS: Thirty male Swiss mice were inoculated with 1x10³ Ehrlich tumor (ET) cells. The mice were divided into three groups (n=10): CG (saline); ACG1 (1.864 mg/Kg of AC) and ACG2 (2.796 mg/Kg of AC), treated every 48 hours from day 3 until day 13. On day 15 the mice were euthanized and the number of viable cells in ascites was determined. In the meantime, ET cells were incubated with AC (0.5, 1.0, 2.0 μg/mL). Cell viability and percentage of growth inhibition (PGI) were checked after one, two, three, four, 18 and 24 hours. RESULTS: There was reduction in volume and number of viable cells in ACG1 and ACG2 compared to CG. In ACG1 one of the animals did not present ascites. In ACG2 two mice did not present ascites and in CG none of the mice present ascites. The action of AC was dose and time dependent and there was no significant difference among the three doses. CONCLUSION: The acetone cyanohydrin promoted reduction of the tumor and also prevented tumor development in 20% of the treated animals.