Cytotoxic activity of Androctonus australis hector venom and its toxic fractions on human lung cancer cell line

Several studies have showed that animal venoms are a source of bioactive compounds that may inhibit the growth of cancer cells, which makes them useful agents for therapeutic applications. Recently, it was established that venom toxins from scorpions induced cytotoxic, antiproliferative and apoptogenic effects on cancer cells. Therefore, the present study aims to investigate the cytotoxic activity of Androctonus australis hector (Aah) scorpion venom and its toxic fractions (FtoxG-50 and F3) on NCI-H358 human lung cancer cells. Methods: The cytotoxic and antiproliferative activities were estimated using MTT assay, lactate dehydrogenase release and clonogenic assays. Apoptosis was evaluated by Hoechst 33258 staining, DNA fragmentation assay and caspase-3 activity. Oxidative stress was analyzed by reactive oxygen species, nitric oxide, malondialdehyde and protein carbonyl levels along with assessment of antioxidant status. In addition, alteration of mitochondrial membrane potential was analyzed by JC1 fluorescent dye. Results: The present findings showed that F3 fraction was more cytotoxic towards NCI-H358 lung cancer cells with an IC50 of 27.05 ± 0.70 g/mL than venom alone (396.60 ± 1.33 g/mL) and its toxic fraction FtoxG-50 (45.86 ± 0.91 g/mL). Nevertheless, F3 fraction was not cytotoxic at these concentrations on normal human lung fibroblast MRC-5 cells. Inhibition of NCI-H358 cell proliferation after F3 fraction exposure occurred mainly by apoptosis as evidenced by damaged nuclei, significant DNA fragmentation level and caspase-3 activation in a dose dependent manner. Moreover, F3 fraction enhanced oxidative and nitrosative stress biomarkers and dissipated mitochondrial membrane potential in lung cancer cells along with significant depletion in cellular enzymatic and non-enzymatic antioxidants. Further, the apoptosis induced by F3 fraction was markedly prevented by the antioxidant N-acetylcysteine (NAC) suggesting the potential mechanism of oxidative stress. Conclusion: These findings suggest that F3 fraction could induce apoptosis in lung cancer cells through involvement of oxidative stress and mitochondrial dysfunction. Hence, these properties make F3 fraction a promising candidate for development of new anticancer agents.

Evaluation of antitumoral and antimicrobial activity of Morinda lcitrifolia L. grown in Southeast Brazil

PURPOSE: To evaluate the antitumor and antimicrobial activity of ethanolic extract of Morinda citrifolia L. fruit cultivated in southeastern Brazil. METHODS: Preparation ethanolic extract of the fruit of Morinda citrifolia L. Culture of melanoma cells B16-F10 for treatment with ethanolic extract of Morinda citrifolia L. fruit to determine cell viability by MTT and determination temporal effect of ethanolic extract fruit on the cell growth B16-F10 for 8 days. Evaluation of antimicrobial activity of ethanolic extract fruit against Staphylococcus aureus and Escherichia coli by determination of Minimum Inhibitory Concentration (MIC). RESULTS: The ethanolic extract of Morinda citrifolia L. fruit (10mg/mL) decreased cellular activity and inhibited 45% the rate of cell proliferation of B16-F10 melanoma treated during period studied. The ethanolic extract of Morinda citrifolia L. fruit demonstrated antimicrobial activity inhibiting the growth of both microorganisms studied. Staphylococcus aureus was less resistant to ethanolic extract of Morinda citrifolia L. fruit than Escherichia coli, 1 mg/mL and 10 mg/mL, respectively. CONCLUSION: What these results indicate that the ethanolic extract of the fruit of Morinda citrifolia L. showed antitumor activity with inhibition of viability and growth of B16-F10 cells and also showed antibacterial activity as induced inhibition of growth of Staphylococcus aureus and Escherichia coli. .

Methanol extract from Vietnamese Caesalpinia sappan induces apoptosis in HeLa cells

BACKGROUND: This study evaluated the cytotoxic activity of extracts from Caesalpinia sappan heartwood against multiple cancer cell lines using an MTT cell viability assay. The cell death though induction of apoptosis was as indicated by DNA fragmentation and caspase-3 enzyme activation. RESULTS: A methanol extract from C. sappan (MECS) showed cytotoxic activity against several of the cancer cell lines. The most potent activity exhibited by the MECS was against HeLa cells with an IC50 value of 26.5 ± 3.2 µg/mL. Treatment of HeLa cells with various MECS concentrations resulted in growth inhibition and induction of apoptosis, as indicated by DNA fragmentation and caspase-3 enzyme activation. CONCLUSION: This study is the first report of the anticancer properties of the heartwood of C. sappan native to Vietnam. Our findings demonstrate that C. sappan heartwood may have beneficial applications in the field of anticancer drug discovery.

Growth Inhibition After Exposure to Transforming Growth Factor-beta1 in Human Bladder Cancer Cell Lines

PURPOSE: Transforming growth factor-beta1 (TGF-beta1) plays a dual role in apoptosis and in proapoptotic responses in the support of survival in a variety of cells. The aim of this study was to determine the function of TGF-beta1 in bladder cancer cells. MATERIALS AND METHODS: The role of TGF-beta1 in bladder cancer cells was examined by observing cell viability by using the tetrazolium dye (MTT) assay after treating the bladder cancer cell lines 253J, 5637, T24, J82, HT1197, and HT1376 with TGF-beta1. Among these cell lines, the 253J and T24 cell lines were coincubated with TGF-beta1 and the pan anti-TGF-beta antibody. Fluorescence-activated cell sorter (FACS) analysis was performed to determine the mechanism involved after TGF-beta1 treatment in 253J cells. RESULTS: All six cell lines showed inhibited cellular growth after TGF-beta1 treatment. Although the T24 and J82 cell lines also showed inhibited cellular growth, the growth inhibition was less than that observed in the other 4 cell lines. The addition of pan anti-TGF-beta antibodies to the culture media restored the growth properties that had been inhibited by TGF-beta1. FACS analysis was performed in the 253J cells and the 253J cells with TGF-beta1. There were no significant differences in the cell cycle between the two treatments. However, there were more apoptotic cells in the TGF-beta1-treated 253J cells. CONCLUSIONS: TGF-beta1 did not stimulate cellular proliferation but was a growth inhibitory factor in bladder cancer cells. However, the pattern of its effects depended on the cell line. TGF-beta1 achieved growth inhibition by enhancing the level of apoptosis.

ATP-Based Chemotherapy Response Assay in Primary or Recurrent Ovarian and Peritoneal Cancer

PURPOSE: To investigate chemosensitivity with an adenosine triphosphate-based chemotherapy response assay in patients with epithelial ovarian or peritoneal cancer according to tumor histology, grade, and disease status. MATERIALS AND METHODS: One hundred specimens were collected during primary or secondary debulking from 67 patients with primary ovarian cancer, 24 patients with recurrent ovarian cancer, 5 patients with primary peritoneal cancer, and 4 patients with recurrent peritoneal cancer; samples were collected between August 2006 and June 2009. Tumor cells were isolated and cultured for 48 hours in media containing chemotherapy. The chemosensitivity index (CI) was calculated as 300 minus the sum of the cell death rate at 0.2x, 1x, and 5x drug concentrations, and the CI values were compared. RESULTS: CI values were obtained from 93 of 100 patients. The most active agents against primary disease were ifosfamide and paclitaxel. For primary serous adenocarcinoma, paclitaxel and irinotecan were the most active, followed by ifosfamide. For clear cell carcinoma, ifosfamide was the most active, followed by paclitaxel and irinotecan. Although not statistically significant, the CIs of cisplatin, carboplatin, paclitaxel, and docetaxel decreased as tumor grade increased. In 14 cases of recurrent disease, paclitaxel was the most active, followed by ifosfamide and cisplatin. CONCLUSION: Ifosfamide and paclitaxel were the most active drugs for primary and recurrent disease. Therefore, we recommend further clinical studies to confirm the efficacy of paclitaxel, ifosfamide, and cisplatin combination chemotherapy for recurrent and primary ovarian cancer.

A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D) models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer.

FTY720 induces apoptosis in B16F10-NEX2 murine melanoma cells, limits metastatic development in vivo, and modulates the immune system

OBJECTIVE: Available chemotherapy presents poor control over the development of metastatic melanoma. FTY720 is a compound already approved by the Food and Drug Administration for the treatment of patients with multiple sclerosis. It has also been observed that FTY720 inhibits tumor growth in vivo (experimental models) and in vitro (animal and human tumor cells). The aim of this study was to evaluate the effects of FTY720 on a metastatic melanoma model and in tumor cell lines. METHODS: We analyzed FTY720 efficacy in vivo in a syngeneic murine metastatic melanoma model, in which we injected tumor cells intravenously into C57BL/6 mice and then treated the mice orally with the compound for 7 days. We also treated mice and human tumor cell lines with FTY720 in vitro, and cell viability and death pathways were analyzed. RESULTS: FTY720 treatment limited metastatic melanoma growth in vivo and promoted a dose-dependent decrease in the viability of murine and human tumor cells in vitro. Melanoma cells treated with FTY720 exhibited characteristics of programmed cell death, reactive oxygen species generation, and increased β-catenin expression. In addition, FTY720 treatment resulted in an immunomodulatory effect in vivo by decreasing the percentage of Foxp3+ cells, without interfering with CD8+ T cells or lymphocyte-producing interferon-gamma. CONCLUSION: Further studies are needed using FTY720 as a monotherapy or in combined therapy, as different types of cancer cells would require a variety of signaling pathways to be extinguished. .

Actividad citotóxica de extractos acuosos de hojas de Trichilia hirta sobre células tumorales humanas / Cytotoxic activity of water extracts of Trichilia hirta leaves on human tumor cells

La planta Trichilia hirta L. (Meliaceae) es utilizada tradicionalmente por pacientes con cáncer como recurso antitumoral. Por ello, los objetivos de este estudio fueron evaluar la actividad citotóxica de extractos acuosos de hojas de Trichilia hirta sobre células tumorales e identificar mediante un tamizaje fitoquímico las principales familias de fitocomponentes presentes en éstos. La actividad citotóxica de los extractos se evaluó sobre células de melanoma humano (SK-mel-3) y adenocarcinoma humano de mama (T-47D). Las células epiteliales de riñón de mono verde Cercopithecus aethiops (Vero) fueron utilizadas como control de células no tumorales. Los resultados mostraron la presencia de triterpenos/esteroides, saponinas, cumarinas, azúcares reductores, fenoles y taninos, flavonoides y carbohidratos/glicósidos en los extractos. Los extractos acuosos de hojas mostraron actividad citotóxica fundamentalmente sobre las células tumorales, lo cual contribuye a explicar la mejoría referida por pacientes con cáncer que consumen estos extractos de forma tradicional

Trichilia hirta L. (Meliaceae) is traditionally used by patients suffering from cancer as an antitumoral resource. Therefore, the objectives of this study were to evaluate the cytotoxic activity of water extracts of Trichilia hirta leaves on tumour cells and identify through a phytochemical screening the principal families of phytocomponents contained in these extracts. The cytotoxic activity of these extracts was also evaluated on human melanoma cells (SK-mel-3) and human breast carcinoma (T-47D). The African green monkey kidney (AGMK) cells Cercopithecus aethiops (Vero) were used as a non-tumour cells control. The results showed the presence of triterpenes/steroids, saponins, coumarins, reductor sugars, phenols and tannins, flavonoids and carbohydrates/glycosides in the extracts. The water leaf extracts showed cytotoxic activity mainly on tumour cells, which contributes to explain the referred recovery by patients suffering form cancer that traditionally consume these extracts

Derivados quiolínicos: compustos sintéticos con propiedades antitumorales sobre líneas de cáncer humano / Quinolinic derivates: synthetic compounds with antitumor proprieties against human line cell cancer

El cáncer es una enfermedad multifactorial caracterizada por anormalidad en el crecimiento celular provocado por factores ambientales y múltiples cambios en la expresión de los genes. El cáncer de mama representa una de las enfermedades que más afecta a la población femenina mundial. Actualmente, se ha mostrado un auge en las investigaciones sobre compuestos extraídos de diversas plantas y origen sintético con posibles agentes antitumorales, entre los que se pueden nombrar los derivados sintéticos de quinolinas. Realizado estudio del grupo sintético indeno (2,1-c) quinolinas y sus efectos citotóxicos sobre líneas de cáncer de mama SKBr3 yMCF-7. La evaluación de la actividad citotóxica de estos compuestos fue determinada a través del método del MTT, se calcularon los valores de concentración inhibitoria, los índices de selectividad y combinaciones con taxanos. Los resultados obtenidos muestran una alta selectividad de los compuestos sintéticos y natural evaluados hacia las líneas celulares MCF-7 y SKBr3 con respecto a las células control (fibroblastos dérmicos humanos); además de una interacción de tipo potenciación en la combinación de un derivado quinolínico y la drogataxano. Con todo esto se puede inferir que solo ciertos compuestos que tuvieron modificaciones químicas realizadas sobre el compuesto patrón del grupo sintético indeno (2,1-c) quinolinas mostraron una inhibición sobre la viabilidad celular sobre las líneas tumorales SKBr3 y MCF-7, conforme aumenta la dosis de los compuestos, además de evidenciar la efectividad de compuestos derivados de quinolinas como moléculas con características antineoplásicas

The cancer is a multifactorial disease characterized by abnormal cell growth caused by environmental factors and for multiple changes in the gene expression. The breast cancer is a disease that affects the female population in worldwide. Currently, there is shown an increment in the research on compounds that we extracted from various plants and synthetic origin with potential antitumor agents, among which can be namedthe synthetic derivatives of the quinolines. A study was performed on an group of synthetic indeno (2,1-c) quinolines and their cytotoxic effects on breast cancer cell lines SKBR3 and MCF-7 was performed. The evaluation of the cytotoxic activity of these compounds was determined through the MTT method; we were calculated inhibitory concentration values, the rates of selectivity and the drug combinations with taxanes. The results show a high selectivity of the natural and the synthetic compounds evaluated towards the cell lines MCF-7 and SKBR3 cells with respect to control (dermal fibroblasts human), besides an enhancement type interaction in the combination of a drug derivative and the quinolinic taxanes. With all this we can infer that only certain compounds had chemical modifications performed on the composite pattern of synthetic group indeno (2, 1-c) quinolines showed an inhibition on cell viability on tumor cell lines SKBR3 and MCF-7, with increasing dose of the compounds, also showing the effectiveness of compounds derived from the quinolines as molecules with the antineoplastic properties and his characteristics

Effects of aspirin on mesenteric lymph nodes of rabbits as basis for its use on lymph nodes metastases / Efeitos da aspirina em linfonodos mesentéricos de coelhos como base para o uso em metástases linfonodais

PURPOSE: To evaluate the effects of aspirin 10% and 20% on mesenteric lymph nodes of rabbits as basis for its use on lymph nodes metastases. METHODS: A total of 20 lymph nodes from 20 rabbits (randomized in four groups) were evaluated. Aspirin solutions 10% (groups A and C) and 20% (groups B and D) were injected into mesenteric lymph nodes of healthy rabbits and had its gross and histological effects evaluated at 24 hours (groups A and B) and at seven days (groups C and D). RESULTS: In the groups A and B evaluated at 24 hours it was observed extensive necrosis and hemorrhage, a significant increase in apoptosis throughout the lymph node with medullary sinuses enlargement and an increase in germinal centers. In the groups C and D evaluated at seven days of solution injection there was also an increase in apoptosis with higher elevation of histiocytes and a significant decrease of necrosis and an increase of giant cells was noticed causing a foreign body chronic inflammation. In all comparisons, there were no differences between the concentrations used (10 and 20%). CONCLUSIONS: The injection of aspirin on lymph nodes caused necrosis and an increase of apoptosis after 24 hours and after seven days of treatment there was regeneration of the lymph nodes, with intense decrease of necrosis and a great elevation of apoptosis. These experimental results support future clinical studies on application of aspirin in the treatment of lymphatic metastases, since the increase of apoptosis is one of the pillars of cancer therapy.

OBJETIVO: Avaliar os efeitos do ácido acetilsalicílico a 10% e 20% em linfonodos mesentéricos de coelhos para posterior embasamento e uso em metástases linfonodais. MÉTODOS: Um total de 20 linfonodos de 20 coelhos (divididos aleatoriamente em quatro grupos) foi avaliado. As soluções de aspirina a 10% (grupos A e C) e 20% (grupos B e D) foram injetadas em linfonodos mesentéricos de coelhos sadios e seus efeitos macroscópicos e histológicos foram avaliados em 24 horas (grupos A e B) e em sete dias (grupos C e D). RESULTADOS: Nos grupos avaliados em 24 horas (A e B) foi verificada intensa necrose e hemorragia, aumento importante de apoptose em todo o linfonodo, com alargamento dos seios medulares e aumento dos centros germinativos. Nos grupos avaliados em sete dias (C e D) também houve aumento da apoptose, com maior elevação de histiócitos e diminuição importante da necrose; a hemorragia foi ausente e aumento de células gigantes foi visualizado, conferindo processo inflamatório crônico do tipo corpo estranho. Não houve diferença entre as concentrações utilizadas (10 e 20%) em nenhuma das comparações. CONCLUSÕES: A injeção de aspirina em linfonodos causou necrose e um aumento de apoptose após 24 horas e após sete dias de tratamento, houve regeneração dos gânglios linfáticos, com diminuição intensa de necrose e grande aumento de apoptose. Uma vez que o aumento de apoptose é um dos pilares dos tratamentos antineoplásicos, estes resultados experimentais embasam eventual aplicação clínica da aspirina no tratamento de metástases linfonodais.

In vitro anti-inflammatory, cytotoxic and antioxidant activities of boesenbergin A, a chalcone isolated from Boesenbergia rotunda (L.) (fingerroot)

The current in vitro study was designed to investigate the anti-inflammatory, cytotoxic and antioxidant activities of boesenbergin A (BA), a chalcone derivative of known structure isolated from Boesenbergia rotunda. Human hepatocellular carcinoma (HepG2), colon adenocarcinoma (HT-29), non-small cell lung cancer (A549), prostate adenocarcinoma (PC3), and normal hepatic cells (WRL-68) were used to evaluate the cytotoxicity of BA using the MTT assay. The antioxidant activity of BA was assessed by the ORAC assay and compared to quercetin as a standard reference antioxidant. ORAC results are reported as the equivalent concentration of Trolox that produces the same level of antioxidant activity as the sample tested at 20 µg/mL. The toxic effect of BA on different cell types, reported as IC50, yielded 20.22 ± 3.15, 10.69 ± 2.64, 20.31 ± 1.34, 94.10 ± 1.19, and 9.324 ± 0.24 µg/mL for A549, PC3, HepG2, HT-29, and WRL-68, respectively. BA displayed considerable antioxidant activity, when the results of ORAC assay were reported as Trolox equivalents. BA (20 µg/mL) and quercetin (5 µg/mL) were equivalent to a Trolox concentration of 11.91 ± 0.23 and 160.32 ± 2.75 µM, respectively. Moreover, the anti-inflammatory activity of BA was significant at 12.5 to 50 µM and without any significant cytotoxicity for the murine macrophage cell line RAW 264.7 at 50 µM. The significant biological activities observed in this study indicated that BA may be one of the agents responsible for the reported biological activities of B. rotunda crude extract.

Avaliação da eficácia antitumoral e toxicidade de lipossomas pH-sensíveis de circulação prolongada contendo cisplatina no tratamento de camundongos portadores de tumor ascítico de Ehrlich

Cisplatina (CDDP) é um dos agentes ativos citotóxicos mais comumente usados notratamento da carcinomatose peritoneal. A inconveniência de seu uso clínico são os efeitoscolaterais sistêmicos, como nefrotoxicidade e mielotoxicidade. Lipossomas pH-sensíveis decirculação prolongada contendo CDDP (SpHL-CDDP) foram desenvolvidos por nosso grupode pesquisa com o objetivo de promover a liberação de CDDP mais próximo do tumor, bemcomo diminuir a toxicidade. O objetivo deste estudo foi avaliar a eficácia antitumoral etoxicidade de SpHL-CDDP, após a administração intraperitoneal em camundongos portadoresde tumor nas fases inicial ou avançada, com uma dose de 12 mg/Kg. A sobrevida foimonitorada e amostras de sangue foram coletadas para análises bioquímicas e hematológicas.Rins, fígado e baço foram removidos para exame histopatológico. As células tumorais foramavaliadas, segundo sua viabilidade e ciclo celular. Os resultados demonstraram que asobrevida de animais tratados com SpHL-CDDP foi maior do que aqueles tratados comCDDP livre

A morte celular causada pelo tratamento com SpHL-CDDP ocorreu através daindução de apoptose com a parada do ciclo celular na fase G0/G1. O tratamento decamundongos que apresentam câncer inicial com ambas as formulações provocou a supressãode granulócitos. Camundongos tratados com CDDP livre apresentaram também diminuição dacontagem de plaquetas, o que sugere alta mielotoxicidade. No modelo de câncer avançado, otratamento com SpHL-CDDP permitiu melhoria da resposta imune. Camundongos portadoresde câncer em estágio inicial e tratados com CDDP livre ou SpHL-CDDP apresentaram menoríndice de uréia/creatinina, em comparação ao grupo controle salina. Esses achados indicamque ambos os tratamentos foram capazes de reduzir o dano renal causado por carcinomatoseperitoneal. A análise microscópica dos rins de camundongos tratados com SpHL-CDDPmostrou alteração morfológica discreta, enquanto necrose tubular foi observada para animaistratados com CDDP livre. Em relação à hepatotoxicidade, nenhuma alteração nos parâmetros

de química clínica foi observada. Estes achados revelam que SpHL-CDDP pode melhorar aeficácia antitumoral e diminuir a toxicidade renal e da medula óssea. A dosagem de VEGF nolíquido ascítico mostrou que ambas as formulações contendo CDDP, em ambos os estágios detratamento, diminuíram a capacidade angiogênica do tumor de Ehrlich, apresentando efeitoantitumoral. O estudo imunológico mostrou que o tratamento com CDDP livre ou SpHLCDDPapresenta um perfil modulado de resposta imune, com diminuição das citocinas próinflamatóriase de citocinas reguladoras. Portanto, estes resultados abrem a possibilidade deuso futuro de SpHL-CDDP para o tratamento da carcinomatose peritoneal

Avaliação da eficácia antitumoral e toxicidade de lipossomas pH-sensíveis de circulação prolongada contendo cisplatina no tratamento de camundongos portadores de tumor ascítico de Ehrlich / Evaluation of antitumor efficacy and toxicity of pH-sensitive liposomes prolonged circulation containing cisplatin in the treatment of mice with Ehrlich ascites tumor

Cisplatina (CDDP) é um dos agentes ativos citotóxicos mais comumente usados notratamento da carcinomatose peritoneal. A inconveniência de seu uso clínico são os efeitoscolaterais sistêmicos, como nefrotoxicidade e mielotoxicidade. Lipossomas pH-sensíveis decirculação prolongada contendo CDDP (SpHL-CDDP) foram desenvolvidos por nosso grupode pesquisa com o objetivo de promover a liberação de CDDP mais próximo do tumor, bemcomo diminuir a toxicidade. O objetivo deste estudo foi avaliar a eficácia antitumoral etoxicidade de SpHL-CDDP, após a administração intraperitoneal em camundongos portadoresde tumor nas fases inicial ou avançada, com uma dose de 12 mg/Kg. A sobrevida foimonitorada e amostras de sangue foram coletadas para análises bioquímicas e hematológicas.Rins, fígado e baço foram removidos para exame histopatológico. As células tumorais foramavaliadas, segundo sua viabilidade e ciclo celular. Os resultados demonstraram que asobrevida de animais tratados com SpHL-CDDP foi maior do que aqueles tratados comCDDP livre. A morte celular causada pelo tratamento com SpHL-CDDP ocorreu através daindução de apoptose com a parada do ciclo celular na fase G0/G1. O tratamento decamundongos que apresentam câncer inicial com ambas as formulações provocou a supressãode granulócitos. Camundongos tratados com CDDP livre apresentaram também diminuição dacontagem de plaquetas, o que sugere alta mielotoxicidade. No modelo de câncer avançado, otratamento com SpHL-CDDP permitiu melhoria da resposta imune. Camundongos portadoresde câncer em estágio inicial e tratados com CDDP livre ou SpHL-CDDP apresentaram menoríndice de uréia/creatinina, em comparação ao grupo controle salina. Esses achados indicamque ambos os tratamentos foram capazes de reduzir o dano renal causado por carcinomatoseperitoneal. A análise microscópica dos rins de camundongos tratados com SpHL-CDDPmostrou alteração morfológica discreta, enquanto necrose tubular foi observada para animaistratados com CDDP livre. Em relação à hepatotoxicidade, nenhuma alteração nos parâmetros de química clínica foi observada. Estes achados revelam que SpHL-CDDP pode melhorar aeficácia antitumoral e diminuir a toxicidade renal e da medula óssea. A dosagem de VEGF nolíquido ascítico mostrou que ambas as formulações contendo CDDP, em ambos os estágios detratamento, diminuíram a capacidade angiogênica do tumor de Ehrlich, apresentando efeitoantitumoral. O estudo imunológico mostrou que o tratamento com CDDP livre ou SpHLCDDPapresenta um perfil modulado de resposta imune, com diminuição das citocinas próinflamatóriase de citocinas reguladoras. Portanto, estes resultados abrem a possibilidade deuso futuro de SpHL-CDDP para o tratamento da carcinomatose peritoneal.

In vitro cytotoxic effects of Brazilian plant extracts on squamous cell carcinoma of the oral cavity

Squamous cell carcinoma (SCC) is the most prevalent cancer of the oral cavity and the fifth most prevalent of all malignancies in males. Many researchers have attempted to develop new treatments that will improve the prognosis of SCC patients. Over 20 percent of the world's biodiversity is located within the Brazilian forests, but little is known about the chemical and/or pharmacological potential of these plants. Certain extracts obtained from Amazon and Atlantic Forest plants have previously been shown to have cytotoxic activity against various cancers. The aim of this study was to screen these extracts for cytotoxic activity against oral SCC cells. The extracts were analyzed for activity against the KB-ADL#12 cell line at various concentrations up to a maximum dose of 100 µg/mL. Comparisons with a control group were performed using one-way ANOVA. Significant cytotoxicity was induced by the extracts obtained from the aerial parts of Picrolemma sprucei (Simaroubaceae), from the leaves and stems of Laetia suaveolens (Salicaceae), from the aerial parts of Abarema auriculata (Fabaceae-Mimosoideae) and from the stem of A. auriculata.

Efficacy of geraniol but not of β-ionone or their combination for the chemoprevention of rat colon carcinogenesis

β-ionone (βI), a cyclic isoprenoid, and geraniol (GO), an acyclic monoterpene, represent a promising class of dietary chemopreventive agents against cancer, whose combination could result in synergistic anticarcinogenic effects. The chemopreventive activities of βI and GO were evaluated individually or in combination during colon carcinogenesis induced by dimethylhydrazine in 48 3-week-old male Wistar rats (12 per group) weighing 40-50 g. Animals were treated for 9 consecutive weeks with βI (16 mg/100 g body weight), GO (25 mg/100 g body weight), βI combined with GO or corn oil (control). Number of total aberrant crypt foci (ACF) and of ACF ≥4 crypts in the distal colon was significantly lower in the GO group (66 ± 13 and 9 ± 2, respectively) compared to control (102 ± 9 and 17 ± 3) and without differences in the βI (91 ± 11 and 14 ± 3) and βI+GO groups (96 ± 5 and 19 ± 2). Apoptosis level, identified by classical apoptosis morphological criteria, in the distal colon was significantly higher in the GO group (1.64 ± 0.06 apoptotic cells/mm²) compared to control (0.91 ± 0.07 apoptotic cells/mm²). The GO group presented a 0.7-fold reduction in Bcl-2 protein expression (Western blot) compared to control. Colonic mucosa concentrations of βI and GO (gas chromatography/mass spectrometry) were higher in the βI and GO groups, respectively, compared to the control and βI+GO groups. Therefore, GO, but not βI, represents a potential chemopreventive agent in colon carcrvpdate=20110329inogenesis. Surprisingly, the combination of isoprenoids does not represent an efficient chemopreventive strategy.

The effect of copaiba balsam on Walker 256 carcinoma inoculated into the vagina and uterine cervix of female rats / Efeito do óleo de copaíba no tumor de Walker 256 inoculado em vagina e colo de útero de ratas fêmeas

Purpose: To verify the copaiba balsam (Copaifera officinalis) effect on Walker 256 carcinoma inoculated into vagina and uterine cervix of rats. Methods: Eighteen female Wistar rats weighing between 180-250g were used, distributed into 2 groups (GCop, GC). On the 1st day of the experiment, 0.3 ml of Walker 256 carcinoma (2x10(6) concentration) was inoculated in both groups; on the 3rd day of the experiment, it was given 4.8 ml/kg of distilled water to the GC group, and 4.8 ml/kg of copaiba balsam to the GCop group. On the 12th day, euthanasia was performed and the tumor was grafted, being weighted and verified its volume. The data were submitted to statistical analysis with ANOVA test. Results: It was observed that copaiba balsam presented a negative inhibitory potential of 70 percent. Conclusion: The copaiba balsam stimulated the tumor growth.

Objetivo: Verificar o efeito do óleo de copaíba da espécie Copaifera officinalis no carcinoma de Walker 256 inoculado em vagina e colo de útero de ratas. Métodos: Foram utilizadas 18 ratas da linhagem Wistar, pesando entre 180-250g, distribuídas em dois grupos (CCop, GC). No 1º dia de experimento, em ambos os grupos foi inoculado 0,3ml de tumor de Walker 256 na concentração de 2x10(6); no 3º dia após essa inoculação, foi iniciada a administração de água destilada na dose de 4,8 ml/kg ao GC, e copaíba na dose de 4,8 ml/kg ao GCop. No 12º dia foi realizada a eutanásia das ratas e ressecado o tumor, sendo este pesado e averiguado seu volume. Os dados obtidos foram submetidos à análise estatística pelo método ANOVA. Resultados: Observou-se que o óleo de copaíba apresentou um potencial inibitório negativo de 70 por cento. Conclusão: O óleo de copaíba estimulou o crescimento tumoral.

Cytotoxic activity of some marine brown algae against cancer cell lines

The aim of this study was to investigate the in vitro cytotoxic activity of total extract of MeOH (70 percent) and partition fractions of hexan, chloroform (CHCL3), ethylacetate (EtOAc) and MeOH-H2O of brown algae species (Sargassum swartzii, Cystoseira myrica, Colpomenia sinuosa) found in the Persian Gulf against in different cell lines including HT-29, Caco-2, T47D, MDA-MB468 and NIH 3T3 cell lines by MTT and AnnexinV-PI assay. The hexan fraction of S. swartzii and C. myrica showed selective cytotoxicity against proliferation of Caco-2 cells (IC50<100 μg/ml) T47D cell line (IC50<100 μg/ml), respectively. S. swartzii and C. myrica were also observed for increasing apoptosis in Caco-2 and T47D cells. Total extract and fractions of C. sinuosa did not show any significant cytotoxicity against the studied cell lines. MDA-MB468 cells were more sensitive to C. myrica than was T47D (IC50 99.9±8.11 vs. 56.50‘± 0.88). This reflects an estrogen receptor independent mechanism for cytotoxicity of the extract. The IC50 of the hexan fraction of C. myrica on T47D parent cells was lower than it was on T47D-TR cells (IC50 99.9±8.11 vs. 143.15 ± 7.80). This finding suggests a role for the MDR-1 in the development of possible future tolerance to the extract.

Individualized Tumor Response Testing for Prediction of Response to Paclitaxel and Cisplatin Chemotherapy in Patients with Advanced Gastric Cancer

The purpose of our study was to determine the most accurate analytic method to define in vitro chemosensitivity, using clinical response as reference standard in prospective clinical trial, and to assess accuracy of adenosine triphosphate-based chemotherapy response assay (ATP-CRA). Forty-eight patients with chemo-naive, histologically confirmed, locally advanced or metastatic gastric cancer were enrolled for the study and were treated with combination chemotherapy of paclitaxel 175 mg/m2 and cisplatin 75 mg/m2 for maximum of six cycles after obtaining specimen for ATP-CRA. We performed the receiver operator characteristic curve analysis using patient responses by WHO criteria and ATP-CRA results to define the method with the highest accuracy. Median progression free survival was 4.2 months (95% confidence interval [CI]: 3.4-5.0) and median overall survival was 11.8 months (95% CI: 9.7-13.8) for all enrolled patients. Chemosensitivity index method yielded highest accuracy of 77.8% by ROC curve analysis, and the specificity, sensitivity, positive and negative predictive values were 95.7%, 46.2%, 85.7%, and 75.9%. In vitro chemosensitive group showed higher response rate (85.7% vs. 24.1%) (P=0.005) compared to chemoresistant group. ATP-CRA could predict clinical response to paclitaxel and cisplatin chemotherapy with high accuracy in advanced gastric cancer patients. Our study supports the use of ATP-CRA in further validation studies.

Heterogeneity of Adenosine Triphosphate-Based Chemotherapy Response Assay in Colorectal Cancer - Secondary Publication

PURPOSE: Adenosine triphosphate-based chemotherapy response assay (ATP-CRA) is a well-documented and validated technology that can individualize chemotherapy for patients with lung, stomach, or breast cancer. This study explored the feasibility of ATP-CRA as a chemosensitivity test in patients with colorectal cancer. MATERIALS AND METHODS: A total of 118 patients who underwent surgical resection for colorectal adenocarcinoma were analyzed for chemosensitivity to 6 anticancer drugs using ATP-CRA. We calculated the cell death rate (CDR) by measuring intracellular ATP levels of drug-exposed cells and untreated controls. RESULTS: Interpretable results were available for 85.5% (118/138) of patients. The mean coefficient of variation for triplicate ATP measurements was 9.2%. The highest CDR was observed in irinotecan (34.0%) and the lowest CDR in etoposide (21.0%). Paclitaxel had the broadest range of CDR (0-86.7%) and 5-FU had the narrowest range of CDR (0-56.8%). The overall highest responsiveness was seen most prevalently in irinotecan (24.7%, 23/93 patients). Irinotecan had the greatest responsiveness in patients with well differentiated and moderately differentiated carcinoma. CONCLUSION: Our study suggests that ATP-CRA could be used to identify patients with colorectal cancer who might benefit from treatment with a specific chemotherapeutic agent.

Impact of Grade, Hormone Receptor, and HER-2 Status in Women with Breast Cancer on Response to Specific Chemotherapeutic Agents by in vitro Adenosine Triphosphate-based Chemotherapy Response Assay

This study was designed to assess whether histological and biological factors of breast cancer can predict chemoresponse to specific agents. Adenosine triphosphate-based chemotherapy response assay (ATP-CRA) was employed to retrieve chemoresponse to 5-fluorouracil (5-FU), doxetaxel, doxorubicin, epirubicin, and paclitaxel in 49 patients. Tumors with high histologic and nuclear grade have higher response rate to doxorubicin (P<0.05) and palitaxel (P<0.05). Estrogen receptor (ER)-negative tumors respond well to doxorubicin (P=0.038), and progesterone receptor (PR)-negative tumors to 5-FU (P=0.039), doxetaxel (P=0.038), doxorubicin (P=0.000), epirubicin (P=0.010), and paclitaxel (P=0.003). Among the breast cancer subtypes determined by ER, PR, and HER-2 immunohistochemical stains, the HER-2+/ER- subtype has a higher response rate to doxorubicin (P=0.008). This in vitro result suggests that the combination of histologic and nuclear grade, hormone receptor, and HER-2 status can be a predictive factor of response to specific chemotherapy agents. Further in vivo study should be followed for clinical trials.