Type 2 diabetes (T2D) is a major public health issue worldwide and it currently affects nearly 26 million people in the United States. It is estimated that one third of Americans will have diabetes by 2050. T2D is a result of chronic insulin resistance and loss of beta-cell mass and function. Both in experimental animals and people, obesity is a leading pathogenic factor for developing insulin resistance, which is always associated with the impairment in energy metabolism, causing increased intracellular fat content in skeletal muscle, liver, fat, as well as pancreatic islets. Constant insulin resistance will progress to T2D when beta-cells are unable to secret adequate amount of insulin to compensate for decreased insulin sensitivity. In the present study, I investigated whether sulforaphane, a natural compound derived from cruciferous vegetables, can prevent high-fat (HF) diet-induced obesity and diabetes in C57BL/6 mice. Dietary intake of sulforaphane (250 mg/kg diet) prevented hyperglycemia and increased insulin sensitivity in HF diet-induced obese mice. Mice treated with sulforaphane had significant lower serum insulin levels (1.93±0.11 μg/dl) as compared to those without treatment (3.09±0.27 μg/dl, P<0.05). In second study, administration of sulforaphane (40 mg/kg body weight daily via gavage) in obese mice enhanced body weight loss and improved insulin sensitivity. Moreover, sulforaphane increased pyruvate oxidation by 28.85% (P<0.05) and enhanced fatty acid oxidation efficiency by 2.2 fold (P<0.05) in primary human muscle cells. These results suggest that sulforaphane may be a naturally occurring insulin-sensitizing agent that is capable of preventing T2D. / Master of Science

Persistent hypertension is associated with a greater incidence of organ damage and conditions such as stroke, heart failure and endstage kidney disease, which results in increased cardiovascular (CV) morbidity and mortality. Among the patients receiving antihypertensive treatment, the level of adequate blood pressure (BP) control (<140/90 mm Hg) is only 30%-50%. These antihypertensive drugs reduce the risk of CV events only by 20% and stroke by 40%, besides causing adverse effects. The spontaneously hypertensive stroke-prone rat (SHRsp) is a good model of essential hypertension. It starts developing hypertension at 5-6 weeks of age which becomes established hypertension at 12-16 weeks. The SHRsp also develops oxidative stress (characterized by low glutathione levels) and inflammation. We have previously shown that consumption of broccoli sprouts (BSp) decreased oxidative stress, inflammation and blood pressure in both male and female SHRsp. BSp contain sulforaphane glucosinolate (SGS), the precursor of the phase 2 protein inducer sulforaphane. BSp low in SGS did not have these effects. Furthermore, BSp did not have any measurable effects on the normal physiology of Sprague Dawley (SD) rats. The objectives of this study were to determine: (1) the importance of a food matrix in causing these beneficial effects, i.e., can one get the same effects by administration of sulforaphane alone?, (2) whether dietary sulforaphane decreases the level of nitrosylated proteins in the kidneys of SHRsp, (3) whether dietary sulforaphane increases the phase 2 enzyme ã-glutamyl-L-cysteine ligase expression (ã-GCS ), (4) whether dietary sulforaphane increases the phase 2 enzyme glutathione reductase (GRed) expression, (5) whether dietary sulforaphane increases the phase 2 enzyme thioredoxin reductase (TrxR1) expression and (6) whether dietary sulforaphane protects renal artery structure. After 1 week of adaptation, the 4 week old female SHRsp and SD rats were divided into four groups and administered daily by gavage: (i) Corn oil (vehicle) alone (Control); (ii) sulforaphane (5 µmol/kg body weight) in corn oil; (iii) sulforaphane (10 µmol/kg body weight) in corn oil; and (iv) sulforaphane (20 µmol/kg body weight) in corn oil. Systolic BP was determined weekly using a standard tail cuff noninvasive BP measurement system (model 29-SSP; Harvard Apparatus, St. Laurent, QC, Canada). The treatment lasted for 15 weeks. At the end of the treatment period, the animals were anesthetized with isoflurane (3%) and the BP was measured by the intra-arterial catheter method using a BP monitor (MK-2000 instrument; Muromachi Kikai Co., Ltd, Tokyo, Japan). Later, the animals were euthanized and perfused with normal saline, and tissues collected for histology, western blot, gene expression study or measurement of reduced glutathione (GSH). The results of the study showed that chronic administration of sulforaphane in SHRsp significantly increased phase 2 proteins (i.e., significantly increased kidney ã-GCS [0.93. ± 0.07 arbitrary unit (AU)] when compared with SHRsp control [0.36 ± 0.05(AU)] , decreased kidney nitrotyrosine (significantly lowered the levels of nitrotyrosine [0.917± 0.16 AU ] when compared with SHRsp control [1.37± 0.2 AU], protected the arterial structure of small resistance vessels in kidneys, and significantly attenuated the increase in blood pressure by 22-43 mm Hg by the end of the study. In conclusion, the results of this thesis demonstrate that: (i) A minimal change in our diet may have a major impact on our health, (ii) The beneficial health effects previously seen with consumption of BSp are due to the conversion of SGS to sulforaphane and (iii) Long term administration of sulforaphane in SHRsp attenuates the increase in BP and vascular alterations

Persistent hypertension is associated with a greater incidence of organ damage and conditions such as stroke, heart failure and endstage kidney disease, which results in increased cardiovascular (CV) morbidity and mortality. Among the patients receiving antihypertensive treatment, the level of adequate blood pressure (BP) control (<140/90 mm Hg) is only 30%-50%. These antihypertensive drugs reduce the risk of CV events only by 20% and stroke by 40%, besides causing adverse effects. The spontaneously hypertensive stroke-prone rat (SHRsp) is a good model of essential hypertension. It starts developing hypertension at 5-6 weeks of age which becomes established hypertension at 12-16 weeks. The SHRsp also develops oxidative stress (characterized by low glutathione levels) and inflammation. We have previously shown that consumption of broccoli sprouts (BSp) decreased oxidative stress, inflammation and blood pressure in both male and female SHRsp. BSp contain sulforaphane glucosinolate (SGS), the precursor of the phase 2 protein inducer sulforaphane. BSp low in SGS did not have these effects. Furthermore, BSp did not have any measurable effects on the normal physiology of Sprague Dawley (SD) rats. The objectives of this study were to determine: (1) the importance of a food matrix in causing these beneficial effects, i.e., can one get the same effects by administration of sulforaphane alone?, (2) whether dietary sulforaphane decreases the level of nitrosylated proteins in the kidneys of SHRsp, (3) whether dietary sulforaphane increases the phase 2 enzyme ã-glutamyl-L-cysteine ligase expression (ã-GCS ), (4) whether dietary sulforaphane increases the phase 2 enzyme glutathione reductase (GRed) expression, (5) whether dietary sulforaphane increases the phase 2 enzyme thioredoxin reductase (TrxR1) expression and (6) whether dietary sulforaphane protects renal artery structure. After 1 week of adaptation, the 4 week old female SHRsp and SD rats were divided into four groups and administered daily by gavage: (i) Corn oil (vehicle) alone (Control); (ii) sulforaphane (5 µmol/kg body weight) in corn oil; (iii) sulforaphane (10 µmol/kg body weight) in corn oil; and (iv) sulforaphane (20 µmol/kg body weight) in corn oil. Systolic BP was determined weekly using a standard tail cuff noninvasive BP measurement system (model 29-SSP; Harvard Apparatus, St. Laurent, QC, Canada). The treatment lasted for 15 weeks. At the end of the treatment period, the animals were anesthetized with isoflurane (3%) and the BP was measured by the intra-arterial catheter method using a BP monitor (MK-2000 instrument; Muromachi Kikai Co., Ltd, Tokyo, Japan). Later, the animals were euthanized and perfused with normal saline, and tissues collected for histology, western blot, gene expression study or measurement of reduced glutathione (GSH). The results of the study showed that chronic administration of sulforaphane in SHRsp significantly increased phase 2 proteins (i.e., significantly increased kidney ã-GCS [0.93. ± 0.07 arbitrary unit (AU)] when compared with SHRsp control [0.36 ± 0.05(AU)] , decreased kidney nitrotyrosine (significantly lowered the levels of nitrotyrosine [0.917± 0.16 AU ] when compared with SHRsp control [1.37± 0.2 AU], protected the arterial structure of small resistance vessels in kidneys, and significantly attenuated the increase in blood pressure by 22-43 mm Hg by the end of the study. In conclusion, the results of this thesis demonstrate that: (i) A minimal change in our diet may have a major impact on our health, (ii) The beneficial health effects previously seen with consumption of BSp are due to the conversion of SGS to sulforaphane and (iii) Long term administration of sulforaphane in SHRsp attenuates the increase in BP and vascular alterations

Le traitement des tumeurs cérébrales représente un défi d'envergure, puisqu'elles sont isolées du reste du corps par la barrière hémato-encéphalique. Il est donc impératif d'accroître nos connaissances sur les cellules composant ces tumeurs. L'une des hypothèses émergentes concerne la présence d'une sous-population spécifique de cellules au sein de la tumeur, les cellules souches cancéreuses (CSC), impliquées dans l'initiation et la récurrence des cancers. Les marqueurs de surfaces des CSC varient largement selon le tissu d'intérêt, mais il semble que CD133, une glycoprotéine membranaire, soit une signature commune à plusieurs CSC. S'il est courant de traiter le cancer en ciblant les cellules cancéreuses elles-mêmes, cibler le microenvironnement dans lequel évolue la tumeur est une avenue thérapeutique prometteuse. En effet, les cellules endothéliales (CE) au sein d'une tumeur sont connues pour avoir des propriétés différentes des CE normales. Ces cellules endothéliales tumorales (CET) peuvent donc constituer une cible supplémentaire pour inhiber sélectivement la croissance tumorale. À l'aide d'une lignée de médulloblastome (DAOY), nous avons identifié et évalué de nouvelles caractéristiques cellulaires et moléculaires impliquées dans la régulation du phénotype invasif associé à la formation de structures comparables aux neurosphères formées par les CSC CD133(+). De plus, nous avons apporté de nouvelles informations concernant les propriétés chimiopréventives du sulforaphane (SFN), un composé naturel retrouvé principalement dans le brocoli, en ciblant spécifiquement les CET cérébrales possiblement impliquées dans la cooption vasculaire. En utilisant la technologie de l'ARN interférant, nous avons montré que MMP-9 et MT1-MMP, deux métalloprotéinases matricielles importantes pour l'invasion cellulaire, les métastases et la résistance à la radiation, avaient un rôle crucial dans la formation par les DAOY de structures apparentées aux neurosphères CD133(+). De plus, nous avons mis en évidence une inhibition presque complète (> 90 %) de la migration des HBMEC (human brain microvascular endothelial cell) activées par un carcinogène, le phorbol 12-myristate 13-acétate (PMA). Ainsi, nos résultats suggèrent que les molécules présentes dans notre diète pourraient cibler les CET cérébrales composant la masse tumorale. En somme, nous proposons que MMP-9 constitue une cible de choix pour le traitement du cancer, que ce soit au niveau du compartiment tumoral ou vasculaire. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Cellule souche cancéreuse (CSC), Cellule endothéliale tumorale (CET), Sulforaphane (SFN), Métalloprotéinase matricielle-9 (MMP-9), Tumeur cérébrale.

Xenobiotics are plant derived compounds metabolized by phase I and II liver enzymes. Phase I enzymes increase, and phase II enzymes decrease, xenobiotic toxicity. Xenobiotics considered were ergotamine, associated with fescue toxicosis, and sulforaphane, a phase II inducer. Hypothesized responses in liver gene expression and enzyme activity due to exposure to these xenobiotics were tested. Polymorphic mice were gavaged with sulforaphane, ergotamine or control over four daily dosing periods (2, 5, 8 and 11 d), with at least 5 mice per treatment. Mice were killed and livers collected 24 h after last dosing. With ergotamine, expression of phase II genes catecholâ Oâ amine methyltransferase 1 (P = 0.009) on d 8, and glutathioneâ Sâ transferase (Gst) mu1 (Gstm1; P = 0.049) on d 11 was increased, and sulfotransferase 5a1 on d 11 decreased (P = 0.02). Sulforaphane increased expression of cytochrome P450 1a2 on d 5 (P = 0.02) and flavin containing monooxygenases 1 on d 11 (P = 0.002), both phase I genes. It also increased expression of a phase II gene transcription factor (P = 0.03) and quinone reductase 02 (P = 0.007) on d 5, and Gstm1 on d 8 (P = 0.04) and d 11 (P = 0.01). Moreover, sulforaphane treated mice had higher (P < 0.05) Gstm1 expression across days. Among enzymes, only sufloraphane treated mice had higher (P < 0.05) Gst activity. The increase in both Gstm1 expression and Gst activity indicate a consistent benefit of sufloraphane on phase II enzyme activity. / Master of Science

A hipertensão arterial pulmonar (HAP) é uma doença grave, caracterizada pelo aumento na resistência vascular pulmonar, elevando a pós-carga imposta ao ventrículo direito. Na tentativa de compensar o aumento da pós-carga, o ventrículo direito desenvolve hipertrofia, que pode evoluir para dilatação e insuficiência, síndrome conhecida como Cor pulmonale. Alterações no estado redox estão presentes na HAP e estão relacionadas ao remodelamento e insuficiência do ventrículo direito. Desta forma, as espécies reativas de oxigênio podem ser alvos terapêuticos interessantes na HAP. O sulforafano é um fitoquímico que vem sido largamente estudado pelo seu potencial de indução de enzimas antioxidantes. Dessa forma, este estudo analisou a influência do tratamento com sulforafano sobre o remodelamento do ventrículo direito e estado redox em um modelo experimental de HAP induzido por monocrotalina. O projeto foi submetido ao Comitê de Ética em Pesquisa com Animais da Universidade Federal do Rio Grande do Sul e aprovado sob o número 26270. Foram utilizados ratos Wistar machos, separados em quatro grupos (n=10-12 animais/grupo): controle (C); Controle + Sulforafano (CS); Monocrotalina (M); Monocrotalina + Sulforafano (MS). A indução da HAP foi realizada por meio de uma dose única de monocrotalina (60 mg/kg – i.p.). O tratamento com sulforafano (2,5 mg/kg i.p.) foi iniciado no 7º dia após a injeção de monocrotalina até 20º dia. Após 21 dias da indução da HAP, os ratos foram anestesiados e foi realizada ecocardiografia, cateterismo e eutanásia dos animais. Os resultados foram analisados através de ANOVA de duas vias e pós-teste de Student Newman Keuls (nível de significância P <0,05). Os resultados serão apresentados na versão completa desta dissertação. / Pulmonary arterial hypertension (PAH) is a serious disease characterized by an increase in pulmonary vascular resistance, increasing afterload imposed on the right ventricle. In an attempt to offset the increased afterload, right ventricular hypertrophy develops, which may progress to dilatation and failure, syndrome known as Cor pulmonale. Alterations in redox state are present in PAH and are related to remodeling and right ventricular failure. Thus, reactive oxygen species may be interesting drug targets in PAH. Sulforaphane is a phytochemical that has been widely studied for its potential induction of antioxidant enzymes. Thus, this study examined the influence of treatment with sulforaphane on the remodeling of the right ventricle and redox state in an experimental model of PAH-induced monocrotaline. The project was submitted to the Comitê de Ética em Pesquisa com Animais of the Universidade Federal do Rio Grande do Sul and approved under number 26270. Male Wistar rats were divided into four groups (n = 10-12 animals/group): Control (C); Control + Sulforaphane (CS); Monocrotaline (M); Sulforaphane + Monocrotaline (MS). Induction of PAH was performed by a single dose of monocrotaline (60 mg/kg - i.p.). The treatment with sulforaphane (2.5 mg/kg i.p.) was initiated on day 7 after injection of monocrotaline to 20 days. After 21 days of induction of PAH, rats were anesthetized and was performed echocardiography, catheterization and euthanasia of the animals. Results were analyzed by two-way ANOVA and Student Newman Keuls post-test (significance level P <0.05). Results will be presented in the full version.

A isquemia seguida de reperfusão está associada com a ativação de uma cascata de eventos danosos intimamente relacionados ao estresse oxidativo. Durante a isquemia, ocorre o aumento da produção de espécies reativas de oxigênio (EROs), que durante a reperfusão, é favorecida pelo restabelecimento da tensão de oxigênio. Uma vez que a utilização de antioxidantes exógenos parece exercer efeitos benéficos apenas nas fases iniciais das doenças cardiovasculares, o sulforafano (SFN) é uma nova estratégia terapêutica que atua estimulando a produção da maquinaria antioxidante endógena. Este composto é um isotiocianato natural encontrado em vegetais crucíferos, como o broto de brócolis, que demonstra ter um efeito cardioprotetor associado a sua ação estimulatória sobre a reserva antioxidante endógena. Poucos estudos até o momento evidenciaram o potencial cardioprotetor do SFN na isquemia e reperfusão miocárdica. O objetivo deste estudo, portanto, foi testar a hipótese de que o pré-tratamento com SFN poderia modular a função ventricular pós-isquêmica, atenuando o estresse oxidativo de corações isolados de ratos submetidos à isquemia e reperfusão. Ratos Wistar machos pesando entre 250 – 300g foram tratados por três dias com SFN (10mg/kg/dia i.p.) ou veículo. Vinte e quatro horas após a última injeção, os ratos foram mortos e seus corações foram retirados rapidamente e submetidos à isquemia global em aparelho do tipo Langendorff. Os corações foram perfundidos com solução Krebs-Henseleit por um período pré-isquêmico de 20 minutos (estabilização), seguido por isquemia global normotérmica de 20 minutos e 20 minutos de reperfusão. Foram avaliados os seguintes parâmetros: frequência cardíaca (FC), pressão sistólica do ventrículo esquerdo (PSVE), pressão diastólica final do ventrículo esquerdo (PDFVE), índice de contratilidade (+dP/dt), índice de relaxamento (-dP/dt) e pressão de perfusão coronariana (PP). Ao final do protocolo, os corações foram pesados e congelados a -80ºC para posteriores análises bioquímicas e moleculares. Foram analisadas a expressão da SOD, CAT, GPx e heme oxigenase-1 (HO-1) e atividade da SOD, CAT, GPx, TrxR, Grx e GST, bem como a produção de EROs totais e lipoperoxidação evidenciada pelas substâncias reativas ao ácido tiobarbitúrico em homogeneizados dos corações pós isquêmicos. Os resultados deste trabalho mostram que o SFN foi capaz de estimular a 6 maquinaria antioxidante intracelular através do aumento significativo de 66% da expressão de ambas as enzimas SOD e HO-1. Ainda, foi capaz de diminuir a produção de EROs totais em 7%. Entretanto, não foi capaz de estimular a expressão de enzimas como a CAT e a GPx. As atividades destas enzimas, bem como das enzimas TrxR, Grx e GST também não apresentaram diferença significativa entre os grupos. Não foram encontradas diferenças entre os grupos também na lipoperoxidação. Quanto à mecânica cardíaca, o SFN não foi capaz de modular a função ventricular pós isquêmica no regime de tratamento utilizado. Como conclusão, o pré tratamento com SFN na dose de 10 mg/kg/dia foi capaz de estimular a expressão de antioxidantes endógenos importantes, tais como a HO-1 e a SOD. Entretanto, o aumento na expressão não repercutiu em suas atividades enzimáticas e, dessa forma, não se observou modificação na função ventricular pós-isquêmica dos corações submetidos à isquemia-reperfusão. / Ischemia followed by reperfusion activates a cascade of injurious events closely related to oxidative stress. During ischemia, generation of reactive oxygen species increases and even more in reperfusion with the reestablishment of oxygen tension. Once exogenous antioxidants seem to exert beneficial effects only in initial phases of cardiovascular diseases, sulforaphane (SFN) is a new therapeutic strategy which acts stimulating endogenous antioxidant machinery. This compound is a natural occurring isothiocyanate, found in cruciferous vegetables as broccoli sprouts, which demonstrates a cardioprotective capacity associated with its capacity of stimulating the antioxidant endogenous reserve. There are few studies until the moment evidenciating the cardioprotective role of SFN on ischemia-reperfusion. The aim of this study was to test the hypothesis that pre-treatment with SFN could modulate the post-ischemic ventricular function, attenuating oxidative stress in isolated hearts submitted to ischemia-reperfusion. Male Wistar rats weighing between 250 – 300g were treated for three days with SFN (10mg/kg/day i.p.) or vehicle. Twenty four hours after the last injection, rats were decapitated and their hearts were rapidly excised and submitted to global ischemia in a Langendorff’s apparatus. Hearts were perfused with a Krebs-Henseleit solution for a pre-ischemic period of 20 minutes (stabilization), followed by normotermic global ischemia of 20 minutes and 20 minutes of reperfusion. The following parameters were evaluated: heart rate (HR), left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), contractility index (+dP/dt), relaxation index (-dP/dt) and coronary perfusion pressure (PP). At the end of protocol, hearts were weighed and frozen at -80ºC for posterior biochemical and molecular analysis. The expression of SOD, CAT, GPx and heme oxygenase-1 (HO-1) and activity of SOD, CAT, GPx, TrxR, Grx and GST, as well as total reactive oxygen species production and lipid peroxidation evidenciated by thiobarbituric reactive substances analysis were performed in homogenates of post-ischemic hearts. The results of the present study show that SFN was capable to stimulate the antioxidant intracellular machinery due to a significant increase of 66% in the expression of both SOD and HO-1 enzymes. Moreover, it was capable to reduce ROS production by about 7%. Although, it was not capable to induce CAT and GPx expression. The activities of these enzymes, as well as TrxR, Grx and GST did not present significant differences between groups. 8 No differences between groups were found in lipid peroxidation. Regarding to mechanical function, SFN was not capable to modulate post-ischemic ventricular function with the treatment regimen used. In summary, pre-treatment with 10 mg/kg/day SFN was capable to stimulate the expression of important endogenous antioxidants, such as HO-1 and SOD. However, the increase on the expression did not reflect in its enzymatic activities and, thereby, no modification on post-ischemic ventricular function of the hearts submitted to ischemia-reperfusion was observed.

Traumatic Brain Injury (TBI) is an increasingly diagnosed constellation of injuries derived from acute mechanical trauma to the brain. With the rise of advanced neuroimaging techniques recent focus has oriented primarily towards the mild-moderate range of TBI which previously was missed diagnostically. Characteristically, these advances have shown increasing areas of micro-hemorrhage in susceptible areas of the brain and to date there are no treatment modalities targeting micro-hemorrhages or their sequelae. This dissertation explores the effects of the resulting heme processing response in the days following injury with a particular focus on inducing early heme clearance from the parenchyma using a rat central fluid percussion injury model in the mild-moderate injury range. Since heme is released ~24-48 hours post-injury and is known to be cytotoxic we observed there may be a critical window for treatment to clear heme before it is spontaneously released and to increase the buffering capacity of the tissue. We targeted heme clearance by using drugs known to increased expression of Nrf2, an upstream transcriptional regulator of the canonical heme processing protein heme oxygenase 1 (HO-1), and tracking expression of HO-1, the iron sequestration/storage proteins Lipocalin 2 (LCN2) and Ferritin (FTL), as well as the activity of matrix metalloproteinases 2 and 9 (MMP2, MMP9). We examined both tissue known to be frankly hemorrhagic (the neocortex) as well as tissue lacking any identifiable bleed (the hippocampus). We demonstrated that using the HO-1 inducers Hemin and Sulforaphane in a single dose paradigm given 1 hour post-injury heme clearance was accelerated in the neocortex with the majority of heme pigment processed by 24 hours post-injury. Further there was significant attenuation of protein expression in HO-1 and ferritin as well as the enzyme activity of MMP2 and MMP9 in both the neocortex and the hippocampus. Behavioral attenuation was also seen in both rotarod and Morris water maze tests. While we intended to target hemorrhagic processing after injury, and indeed demonstrated improved clearance of heme from post-injury hemorrhagic regions of the brain, in both tissues studied we observed remarkably similar responses to the drugs utilized in protein expression, enzyme activity, and behavioral improvement which may suggest a globally improved pathologic state or that there are unidentified pathologic micro-hemorrhages or leaky vessels which extend further into the brain parenchyma than currently identified.

Objective: To evaluate the anti-tumor and histone deacetylase (HDAC) inhibitory activity of the dietary isothiocyanate, sulforaphane (SFN) in the paediatric cancer,neuroblastoma (NB). Materials and Methods: NB cell line (NUB-7), fibroblasts (FLF; negative control) and MCF-7 (positive control), were treated with SFN for up to 7 days and effects on growth, cytotoxicity, differentiation and tumorigenicity assessed. HDAC inhibition was determined by histone (H3/ H4) acetylation. Results: 10 μM SFN significantly decreased in vitro growth and survival of NUB-7 to 10.22 ± 0.71% (p < 0.001) with no significant effect on FLF. SFN induced G1, G2 and S phase cell cycle arrests and stimulated H3/H4 histone acetylation. SFN markedly decreased NUB-7 clonogenicity and tumorigenicity in vivo. Conclusion: Results suggest that low dose SFN reduces proliferation, survival and tumorigenicity of NB NUB-7. As a dietary factor of negligible intrinsic toxicity SFN is a promising therapeutic agent for the treatment of NB.

Objective: To evaluate the anti-tumor and histone deacetylase (HDAC) inhibitory activity of the dietary isothiocyanate, sulforaphane (SFN) in the paediatric cancer,neuroblastoma (NB). Materials and Methods: NB cell line (NUB-7), fibroblasts (FLF; negative control) and MCF-7 (positive control), were treated with SFN for up to 7 days and effects on growth, cytotoxicity, differentiation and tumorigenicity assessed. HDAC inhibition was determined by histone (H3/ H4) acetylation. Results: 10 μM SFN significantly decreased in vitro growth and survival of NUB-7 to 10.22 ± 0.71% (p < 0.001) with no significant effect on FLF. SFN induced G1, G2 and S phase cell cycle arrests and stimulated H3/H4 histone acetylation. SFN markedly decreased NUB-7 clonogenicity and tumorigenicity in vivo. Conclusion: Results suggest that low dose SFN reduces proliferation, survival and tumorigenicity of NB NUB-7. As a dietary factor of negligible intrinsic toxicity SFN is a promising therapeutic agent for the treatment of NB.