Efeito quimiopreventivo da Beta-ionona na hepatocarcinogênese associada à fibrose hepática / Chemopreventive effect of Beta-ionone in hepatocarcinogenesis associated with hepatic fibrosis

O carcinoma hepatocelular (HCC) é a neoplasia primária mais frequente que acomete o fígado, a quarta principal causa de morte relacionada ao câncer e apresenta mau prognóstico. A fibrose hepática está presente em grande parte dos casos de HCC e é um dos principais fatores de risco para esta afecção. Segundo estudos prévios do grupo, a ß-ionona (BI), presente em uvas e aromatizantes de vinho, apresenta potencial quimiopreventivo na hepatocarcinogênese principalmente por reduzir o número e tamanho de lesão pré neoplásica (LPN) e inibir a proliferação celular. No entanto, até o presente não foram identificados na literatura estudos que investigaram o efeito deste isoprenóide no processo fibrótico e na hepatocarcinogênese a ele associada. Desta forma, este estudo pretendeu investigar o potencial efeito quimiopreventivo da BI na hepatocarcinogênese associada à fibrose hepática. Para tanto, ratos machos Wistar foram tratados com óleo de milho (OM) [0,25 ml / 100 g de peso corporal (p.c.); Grupo de OM] ou BI (16mg / 100g p.c.; Grupo BI) durante 18 semanas. A partir da 2ª semana, todos os animais receberam uma dose intraperitoneal de dietilnitrosamina (DEN - 50 mg / Kg p.c.) uma vez por semana até a 16ª semana. Os animais foram eutanasiados em diferentes períodos do protocolo experimental: na 10a semana (grupos OMP1 e BIP1), na 14a semana (grupos OMP2 e BIP2) e na 18ª semana (grupos OMP3 e BIP3). O isoprenóide demonstrou, de maneira inédita na literatura, inibir o desenvolvimento da fibrose hepática em diferentes estágios da hepatocarcinogênese (pontos 1, 2 e 3) por reduzir (p < 0,05) a porcentagem de área marcada para picrosirius. Além disso, BI reduziu a porcentagem de área positiva para α- SMA (p < 0,05) e as concentrações de hidroxiprolina (p < 0,05) no ponto 2. Foi observada ação quimiopreventiva da BI nas fases iniciais da hepatocarcinogênese (pontos 1 e 2) mesmo em modelo associada a fibrose por reduzir (p < 0,05) o número e porcentagem de área do corte ocupada por LPN GSTP positivas. Este efeito não foi observado em fase mais avançada da hepatocarcinogênese (ponto 3). Corroborando este dado não foram observadas diferenças em relação ao número de tumores (p>=0,05) avaliados por imageamento e por análise histopatológica. No entanto, quando comparados ao seu controle (OMP3), os animais do grupo BIP3 apresentaram menor mortalidade e menor incidência (p < 0,05) de HCC high, considerado um tipo mais agressivo de HCC, sugerindo que este composto possa atuar na agressividade das células tumorais. O grupo BIP2 demonstrou ainda menor proliferação celular (p < 0,05) quando comparado ao grupo OMP2. Assim foram avaliadas as vias de proliferação celular PI3K/AKT e MAPK/ERK, bem como as proteínas p21 e p53, relacionadas a progressão do ciclo celular. Não foram observadas(p≥0,05) alterações nestas vias por parte do isoprenóide. O presente estudo demonstrou ação protetora da BI no desenvolvimento de fibrose, bem como na hepatocarcinogênese a ela associada. Contudo, são necessárias análises complementares para elucidar mecanismos pelos quais a BI atua na carcinigênese hepática associada à fibrose

Hepatocellular carcinoma (HCC) is the primary liver cancer, the fourth leading cause of death related to cancer and presents a poor prognosis. Hepatic fibrosis is present in most cases of HCC and represents one of the main risk factors for this condition. According to previous studies of the group, ß-ionone (BI), present in grapes and wine flavorings, has a potential chemopreventive in hepatocarcinogenesis mainly by reducing the number and size of preneoplastic lesions (LPN) and inhibiting cell proliferation. However, to date, no studies have been identified in the literature that investigated the effect of this isoprenoid on the fibrotic process and in its association with hepatocarcinogenesis. Thus, this study aimed to investigate the potential chemopreventive effect of BI in hepatocarcinogenesis associated with hepatic fibrosis. Male Wistar rats were treated with corn oil (OM) [0.25 ml / 100 g body weight (b.w.); OM] or BI group (16mg / 100g b.w; BI group) for 18 weeks. From week 2, all animals received an intraperitoneal dose of diethylnitrosamine (DEN - 50 mg / kg b.w.) once in a week until week 16. The animals were euthanized at different periods of the experimental protocol: at week 10 (groups OMP1 and BIP1), at week 14 (groups OMP2 and BIP2) and week 18 (groups OMP3 and BIP3). The isoprenoid, for the first time in the literature, shown to inhibit the development of liver fibrosis at different stages of hepatocarcinogenesis (points 1, 2 and 3) by reducing (p <0.05) the percentage of the area labeled for picrosirius. Also, BI reduced the percentage of α-SMA positive area (p <0.05) and hydroxyproline concentrations (p <0.05) at point 2. BI chemopreventive action was observed in the early stages of hepatocarcinogenesis (point 1 and 2) even in a model associated with fibrosis for reducing (p <0.05) the number and percentage of the liver section area occupied by GSTP positive LPN. This effect was not observed at a later stage of hepatocarcinogenesis (point 3). Corroborating this data, no differences were observed regarding the number of tumors (p>=0.05) evaluated by imaging and histopathological analysis. However, when compared to its control (OMP3), animals from the BIP3 group had lower mortality and lower incidence (p<0.05) of HCC high, considered a more aggressive type of HCC, suggesting that this compound may act in aggressiveness of tumor cells. The BIP2 group also showed lower cell proliferation (p <0.05) when compared to the OMP2 group. Thus, PI3K / AKT and MAPK / ERK cell proliferation pathways were evaluated, as well as p21 and p53 proteins, related to cell cycle progression. No changes were observed in these pathways by the isoprenoid (p≥0.05). The present study demonstrated the protective action of BI in the development of fibrosis, as well as its association with hepatocarcinogenesis. However, further analysis is needed to elucidate mechanisms by which BI acts on fibrosis-associated liver carcinogenesis

Ethanol extract of Phellinus merrillii protects against diethylnitrosamine- and 2-acetylaminofluorene-induced hepatocarcinogenesis in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study whether the ethanol extract of Phellinus merrillii (EPM) has chemopreventive potential against liver carcinogenesis.</p><p><b>METHODS</b>Thirty male Spraque-Dawley rats were randomly divided into control group, EPM control group, hepatocarcinoma control group, low-dose EPM group and high-dose EPM group, 6 in each group. Using the Solt and Farber protocol in a rat model of hepatocarcinogenesis, the chemopreventive effect of EPM on diethylnitrosamine (DEN)-initiated, 2-acetylaminofluorene (2-AAF) and partial hepatectomy (PH)-promoted liver carcinogenesis in rats was evaluated. Basic pathophysiological and histological examinations, together with the serum levels of glutamic oxaloacetic transaminase (sGOT), glutamic pyruvic transaminase (sGPT) and gamma-glutamyl transpeptidase (γ-GT) were measured.</p><p><b>RESULTS</b>Treatment of EPM at the concentration of 2 g/kg body weight in the diet for 8 weeks clearly prevented the development of carcinogenesis and reduced the levels of sGOT, sGPT, and serum γ-GT of rats as compared with the hepatocarcinoma control group (P<0.05 or P<0.01). These phenotypes were accompanied by a significant increase in natural killer cell activity.</p><p><b>CONCLUSION</b>EPM showed a strong liver preventive effect against DEN+2-AAF+PH-induced hepatocarcinogenesis in a rat model.</p>

Bee Honey Modulates the Oxidant-Antioxidant Imbalance in Diethyl nitrosamine-Initiated Rat Hepatocellular Carcinoma.

Oxidative stress reflects the mechanism that contributes to initiation and progression of hepatic injury in a variety of liver disturbance. From here, there is a great demand for the expansion of agents with a potent antioxidant effect. The aim of this work is to approximate the efficiency of bee honey as a hepatoprotective and an antioxidant agent versus diethyl nitrosamine (DEN) motivate hepatocellular damage. The single intrapritoneal (IP) management of diethyl nitrosamine (50mg/kg followed by 2ml/kg CCl4) to rats, referred for the histopathological examination of liver sections of rats after induction and before treatment with honey showed that many well differentiated tumor cells were formed in the liver of rats also, the examined sections showed disorganization of hepatic lobular architecture and obvious cellular damage. A significant lift in the enzymatic activity of liver functions (AST, ALT, ALP), and gamma glutamyltransferase (GGT) which is a signal of hepatocellular damage. DEN stimulates oxidative stress, which was assured by increase lipid peroxidation level and hindrance in antioxidant enzymes (SOD, CAT, GPx, and GST) activities in the liver. The position of non-enzymatic antioxidants comparable reduced glutathione (GSH) was likewise set up to be slimmed down significantly in DEN inoculated rats. Also, we have studied the underlying mechanism and /or (s) of the therapeutic role of bee honey as hepatocarcinogenesis remediation through investigation the inflammatory biomarkers; α-fetoprotein (AFP) and α-fucosidase (AFU). The current results clearly showed that bee honey demonstrates good ameliorative and antioxidant capacity toward diethyl nitrosamine induced hepatocellular damage in rats.

Potencial quimiopreventivo de lipídios estruturados obtidos por interesterificação da tributirina com o óleo de linhaça na hepatocarcinogênese / Chemopreventive potential of structured lipids obtained by interesterification of tributyrin with flaxseed oil in hepatocarcinogenesis

O carcinoma hepatocelular (HCC) apresenta mau prognóstico o que torna importante sua quimioprevenção. Nesse sentido, a tributirina (TB), um inibidor de desacetilases de histonas (HDACi), mostrou-se um quimiopreventivo promissor da hepatocarcinogênese. Avaliaram-se aqui efeitos quimiopreventivos de lipídios estruturados (EST) obtidos por interesterificação enzimática a partir da TB com o óleo de linhaça (LIN). Ratos foram tratados com EST (grupo EST; 165 mg/100g peso corpóreo [p.c]), TB (grupo TB; 200 mg/100g p.c), LIN (grupo LIN; 133 mg/100g p.c), mistura de TB com LIN (grupo LIN; 165 mg/100g p.c) ou maltodextrina (MD) (grupo MD; controle isocalórico; 300 mg/100g p.c) diariamente durante 8 semanas consecutivas por gavagem. Duas semanas após início dos tratamentos, os animais foram submetidos ao modelo de hepatocarcinogênese do "hepatócito resistente" (RH). Os grupos EST e TB apresentaram atividade quimiopreventiva bloqueadora e supressora, respectivamente, da hepatocarcinogênese. TB induziu a apoptose, ao contrário dos EST. O tratamento com TB resultou na acetilação e trimetilação da H3K9 e H3K27, enquanto EST atuaram somente na trimetilação das mesmas. Quando analisada a expressão de genes envolvidos com modificações em histonas, EST e TB reduziram a expressão de Ezh2 e de Hdac4. Por outro lado, somente os EST aumentaram a expressão de Hdac6. Tal efeito por parte dos EST merece ser mais investigado, uma vez que esta desacetilase vem sendo sugerida como alvo potencial para o desenvolvimento de fármacos. Em conclusão, a atividade quimiopreventiva de EST e da TB envolve na hepatocarcinogênese experimental mecanismos epigenéticos que podem ou não ser distintos

Hepatocellular carcinoma (HCC) has a poor prognosis, which makes its chemoprevention important. Tributyrin (TB), which is a histone deacetylase inhibitor (HDACi), is a promising chemopreventive agent of hepatocarcinogenesis. The chemopreventive effects of structured lipids (STLs) that were obtained by the enzymatic interesterification of TB with flaxseed oil (FSO) were evaluated in the present study. Rats were treated with STLs (STL group, 165 mg/100 g body weight (bw)), TB (TB group, 200 mg/100 g bw), FSO (FSO group, 133 mg/100 g bw), TB mixed with FSO (BLD group, 165 mg/100g bw) or maltodextrin (MD) (MD group; isocaloric control; 300 mg/100 g bw) daily for eight consecutive weeks by gavage. Two weeks after the initiation of treatment, the animals were subjected to the resistant hepatocyte hepatocarcinogenesis model (RH). The STL and TB groups developed blocker and suppressive chemopreventive activity against hepatocarcinogenesis, respectively. TB treatment induced apoptosis, unlike the STL treatment. Additionally, TB treatment resulted in the acetylation and trimethylation of H3K9 and H3K27, whereas the STLs acted only in the trimethylation of these histones. When analyzing the expression of genes involved in histone modifications, the STLs and TB reduced enhancer of zeste homolog 2 (Ezh2) and histone deacetylase 4 (Hdac4) gene expression. Conversely, only the STLs increased Hdac6 gene expression. This effect of the STLs warrants further investigation because this deacetylase has been suggested as a potential drug development target. In conclusion, the chemopreventive activities of the STLs and TB in experimental hepatocarcinogenesis involve epigenetic mechanisms that may be distinct

Relationship between hepatitis B virus X protein and hepatic cellular cancer / 药学学报

Hepatic cellular cancer (HCC) is one of the most common cancers in the world, which is a serious threat to human health and life quality. More than 700 000 people die of HCC each year on average, and its incidence increases in many countries. Chronic hepatitis B virus (HBV) infection has been identified as a dominant risk factor for HCC. The pathogenesis of HBV-induced hepatocarcinogenesis is, however, incom-pletely understood. Evidence currently available supports a key role of the HBV X protein (HBx) in the cancer transformation and malignant tumor metastasis. HBx is a multifunctional regulator that may cooperate with the host factors to exert its effects on transcription, signal transduction, cell cycle progression, apoptosis, protein degradation, expression of oncogene and anti-oncogene. This review presents the current knowledge in the molecular pathogenesis of HBx in the induction of HCC.

Regulatory effect of a Chinese medicine prescription Hu Qi Shan on the telomerase activity in hepatic precancerous lesions in rats / 中国比较医学杂志

Objective To explore the possible mechanism of action of telomerase in hepatic precancerous lesions, and the regulatory effect of a Chinese medicine prescription HU Qi Shan ( HQS) and its principal drug mistletoe alkali on the telomerase activity in rat liver tissues.Methods Rat model of hepatic precancerous lesions was established by Solt-Farber two-step protocol.The model rats were randomly divided into 5 groups, including the model group, high-dose HQS [8 g/(kg· d)] group, low-dose HQS [4 g/(kg· d)] group, and mistletoe alkali[8 mg/(kg· d)] group.γ-Glutamy-transpeptidase (γ-GT) was analyzed by immunohistochemistry.AFP was detected by immunofluorescence technique.The telomerase activity was detected using a quantitative telomerase detection kit.The expression of NF-κB P65 was detected by immunohistochemistry.The cytoplasmic protein IκB-αwas detected by western blotting.Results After treated with HQS and mistletoe alkali, the areas ofγ-GT-positive foci and number of AFP-positive cells in the liver tissus were significantly decreased than those of the model group ( P<0.05 for both) , the telomerase activity was decreased, the number of NF-κB P65-positive cells was also decreased ( P <0.05 ) , whereas the intracytoplasmic expression of IκB-αproteins was significantly increased ( P <0.05 ) .Conclusions HQS and mistletoe alkali can suppress the telomerase activity.Its possible mechanism may be through inhibition of the over-expressed apoptosis-related genes such as NF-κB P65 and increase the expression of IκB-αdecreasing the telomerase activity.

Pathology-MRI Correlation of Hepatocarcinogenesis: Recent Update

Understanding the important alterations during hepatocarcinogenesis as well as the characteristic magnetic resonance imaging (MRI) and histopathological features will be helpful for managing patients with chronic liver disease and hepatocellular carcinoma. Recent advances in MRI techniques, such as fat/iron quantification, diffusion-weighted images, and gadoxetic acid-enhanced MRI, have greatly enhanced our understanding of hepatocarcinogenesis.

Tratamiento oncológico / Chemotherapy

Los esquemas de quimioterapia convencionales no han demostrado ningún beneficio en el tratamiento del paciente con carcinoma hepatocelular (CHC), las tasas de respuesta han sido muy bajas y no se ha observado ningún beneficio en la sobrevida cuando se usan agentes únicos o en combinación de quimioterapia. El advenimiento de nuevas moléculas de acción en sitios más específicos ha brindado una mejor esperanza en la sobrevida con menores efectos adversos.

Conventional chemotherapy has shown no benefit for the treatment of patients with hepatocellular carcinoma (HCC). Response rates have been very low and no survival benefits have been observed for any single agent or combination chemotherapy regime. Nevertheless, the advent of new molecules which act on specific sites has given more hope for better survival with fewer adverse effects from chemotherapy.

Expression of CD44 in mouse diethylnitrosamine (DEN)-induced hepatic tumors as well as in DEN-treated embryo stem cells and derived hepatic lineage cells / 동물의과학연구지

To clarify the role of stem cells in hepatocarcinogenesis, CD44 expression was investigated in mouse livers as well as embryonic cell lineages treated with diethylnitrosamine (DEN). Liver tumors induced by DEN were analyzed by immunohistochemisty for CD44. Liver tissues were sampled at 6, 24, and 48 hr after treatment with saline or DEN. Mouse embryonic stem cells (ESCs), hepatic progenitor cells (HPCs), and hepatocyte like cells (HCs), representing 0, 22, and 40 days of differentiation, respectively, were treated with DEN at four doses (0, 1, 5, and 15 mM, respectively) for 24 hr, after which CD44 expression levels were examined by relative quantitative real-time PCR. CD44 expression was weakly detected in tumor cells as well as in some hepatocytes surrounding the tumor cells. However, CD44 expression was not detected in liver tissue treated with DEN at early time points. The CD44 mRNA expression level was significantly different among cells treated with 5 mM DEN at day 22 (P<0.01) as well as 1, 5, and 15 mM DEN at day 40 (P<0.01) compared with control. Taken together, CD44 expression slightly increased in mouse DEN-induced tumors. Furthermore, expression of CD44 in embryonic cell lineages treated with various doses of DEN significantly differed among embryo stem cells and derived hepatic lineage cells. This suggests that CD44 expression may be modulated in the progeny of stem cells during their differentiation toward hepatocytes, and its expression may increase in the tumor stage but not during early carcinogenesis.

Evaluation of the efficacy of hydrated sodium aluminosilicate in the prevention of aflatoxin-induced hepatic cancer in rainbow trout / Avaliação da eficácia do aluminosilicato de sódio hidratado na prevenção de câncer hepático induzido por aflatoxina em truta arco-íris

The use of aluminum silicates for decontaminating animal feed containing aflatoxins has yielded encouraging results in chicken and turkey poults. In contrast, very few studies have tested these substances in aquaculture. In this work, we investigated the efficacy of a trout diet containing 0.5 percent hydrated sodium aluminosilicate (HSAS) in protecting against contamination with aflatoxin B1. Trout were reared on these diets for one year and the experimental groups were examined monthly for hepatic presumptive preneoplastic and neoplastic lesions. Regardless of the presence of HSAS, all of the fish that received aflatoxin in their diet have shown hepatic lesions indicative of a carcinogenic process, presenting also the development of cancer in some fish. The concentration of HSAS used in this study was ineffective in preventing the onset of hepatic lesions induced by aflatoxin B1 in rainbow trout.

Resultados encorajadores têm sido observados com o emprego de aluminosilicatos para descontaminação de ração contendo aflatoxina destinada à avicultura. No entanto, raros estudos têm sido destinados a testar essa substância em aqüicultura. Assim, no presente trabalho, foi investigada a eficácia do aluminosilicato de sódio hidratado (HSAS, 0,5 por cento) em proteger a truta arco-íris dos danos hepáticos causados por ração contaminada com aflatoxina B1. Os grupos experimentais foram alimentados com suas respectivas dietas durante 12 meses, mensalmente cada grupo foi amostrado para análises anatomopatológica e histopatológica para verificação da presença de lesões hepáticas. Apesar da presença de HSAS, todos os grupos que receberam dieta contaminada com aflatoxina B1 apresentaram lesões hepáticas indicativas do processo carcinogênico, sendo que exemplares alimentados com essas dietas por mais longo prazo apresentaram hepatocarcinoma ou colangiocarcinoma. Estes resultados indicaram que a concentração de HSAS usada no presente estudo não foi efetiva em prevenir o desenvolvimento de lesões hepáticas induzidas por aflatoxina B1 em truta arco-íris.

Water extracts of cabbage and kale inhibit ex vivo H2O2-induced DNA damage but not rat hepatocarcinogenesis

The chemopreventive potential of water extracts of the Brassica vegetables cabbage and kale was evaluated by administering their aqueous extracts in drinking water ad libitum to Wistar rats submitted to Ito’s hepatocarcinogenesis model (CB group and K group, respectively - 14 rats per group). Animals submitted to this same model and treated with water were used as controls (W group - 15 rats). Treatment with the vegetable extracts did not inhibit (P > 0.05) placental glutathione S-transferase-positive preneoplastic lesions (PNL). The number of apoptotic bodies did not differ (P > 0.05) among the experimental groups. Ex vivo hydrogen peroxide treatment of rat livers resulted in lower (P < 0.05) DNA strand breakage in cabbage- (107.6 ± 7.8 µm) and kale- (110.8 ± 10.0 µm) treated animals compared with control (120.9 ± 12.7 µm), as evaluated by the single cell gel (comet) assay. Treatment with cabbage (2 ± 0.3 µg/g) or kale (4 ± 0.2 µg/g) resulted in increased (P < 0.05) hepatic lutein concentration compared with control (0.5 ± 0.07 µg/g). Despite the absence of inhibitory effects of cabbage and kale aqueous extracts on PNL, these Brassica vegetables presented protection against DNA damage, an effect possibly related to increased hepatic lutein concentrations. However, it must be pointed out that the cause-effect relationship between lutein levels and protection is hypothetical and remains to be demonstrated.

Atividade quimiopreventiva do ácido fólico quando suplementado continuamente durante as etapas iniciais da hepatocarcinogênese em ratos / Chemoprevention of early rat hepatocarcinogenesis with folic acid supple

A ingestão de folato é inversamente associada com o risco de diversos cânceres. Apesar da deficiência dessa vitamina ser classicamente considerada fator de risco para câncer de fígado, não existem estudos avaliando o efeito da suplementação com ácido fólico (AF) durante as etapas iniciais da hepatocarcinogênese. Assim, o objetivo do presente estudo foi avaliar o efeito da suplementação com AF continuamente durante as etapas de iniciação e seleção/promoção da hepatocarcinogênese em ratos. Os animais receberam diariamente 0,08 mg (grupo AF8) ou 0,16 mg (grupo AF16) de AF/100 g de peso corpóreo ou água (grupo controle [GC]). Após duas semanas de tratamento, todos os animais foram submetidos ao modelo de hepatocarcinogênese do “Hepatócito Resistente” (iniciação com dietilnitrosamina, seleção/promoção com 2-acetilaminofluoreno e hepatectomia parcial a 70%). A eutanásia dos animais ocorreu após 8 semanas de tratamento. Quando comparado ao GC, o grupo AF16, mas não o AF8, apresentou menores nódulos macroscópicos (p<0,05), menor (p<0,05) número de lesões pré-neoplásicas (LPN) persistentes, maior (p<0,05) número de LPN em remodelação, menor (p<0,05) proliferação celular nas LPN persistentes, menos (p<0,05) danos no DNA hepático e tendência (p<0,10) a apresentar menor expressão de c-myc em LPN microdissecadas. Não foram observadas diferenças significativas (p>0,05) entre os grupos experimentais com relação à indução de apoptose nas LPN persistentes e em remodelação bem como no padrão de metilação global do DNA em LPN microdissecadas. Em resumo, a suplementação com AF durante as etapas iniciais da hepatocarcinogênese resultou em atividade quimiopreventiva de forma dose-efeito. Alteração no fenótipo das LPN, inibição de danos no DNA hepático e da expressão de c-myc representam relevantes efeitos celulares e moleculares dessa vitamina.

Dietary intake of folate is inversely associated with the risk of several malignancies. Although folate deficiency is associated with liver cancer, there is no data on folic acid (FA) supplementation during hepatocarcinogenesis. The aim of the present study was to evaluate the effect of FA supplementation during early hepatocarcinogenesis. Rats receiving daily 0.08 mg (FA8 group) or 0.16 mg (FA16 group) of FA/100 g body weight or water (CO group, controls) were used. After a 2 week-treatment, all animals were submitted to the resistant hepatocyte model of hepatocarcinogenesis (initiation with diethylnitrosamine, selection/promotion with 2-acetylaminofluorene and partial hepatectomy). All animals were euthanized after 8 weeks of treatment. When compared to CO group, FA16 group, but not FA8 group, presented: smaller (p < 0.05) macroscopic nodules; reduced (p < 0.05) number of persistent and increased (p < 0.05) number of remodeling preneoplastic lesions (PNL); reduced (p < 0.05) cell proliferation in persistent PNL; decreased (p < 0.05) hepatic DNA damage; and a tendency (p < 0.10) of decreased c-myc expression in microdissected PNL. No differences (p > 0.05) were observed between CO, FA8 and FA16 groups regarding apoptosis in both persistent and remodeling PNL, and global DNA methylation pattern in microdissected PNL. In conclusion, FA supplementation during early hepatocarcinogenesis resulted in a dose-response chemopreventive activity. Reversion of PNL phenotype and inhibition of DNA damage and of c-myc expression represent relevant FA cellular and molecular effects.

Homeopathic drugs Natrum sulphuricum and Carcinosin prevent azo dye-induced hepatocarcinogenesis in mice.

The study was undertaken to examine whether Carcinosin-200 (Car-200) could provide additional ameliorative effect, if used intermittently with Natrum sulphuricum-30 (Nat Sulph-30) against hepatocarcinogenesis induced by chronic feeding of p-dimethylaminoazobenzene (p-DAB) and phenobarbital (PB) in mice (Mus musculus). Mice were randomly divided into seven sub-groups: (i) normal untreated; (ii) normal + succussed alcohol; (iii) p-DAB (0.06%) + PB (0.05%); (iv) p-DAB + PB + succussed alcohol, (v) p-DAB + PB + Nat Sulph-30, (vi) p-DAB + PB + Car-200, and (vii) p-DAB + PB + Nat Sulph-30 + Car-200. They were sacrificed at 30, 60, 90 and 120 days for assessment of genotoxicity through cytogenetical end-points like chromosome aberrations, micronuclei, mitotic index and sperm head anomaly and cytotoxicity through assay of widely accepted biomarkers and pathophysiological parameters. Additionally, electron microscopic studies and gelatin zymography for matrix metalloproteinases (MMPs) were conducted in liver at 90 and 120 days. Results showed that administration of Nat Sulph-30 alone and in combination with Car-200 reduced the liver tumors with positive ultra-structural changes and in MMPs expression, genotoxic parameters, lipid peroxidation, -glutamyl transferase, lactate dehydrogenase, blood glucose, bilirubin, creatinine, urea and increased GSH, glucose-6-phosphate dehydrogenase, superoxide dismutase, catalase, glutathione reductase activities and hemoglobin, cholesterol, and albumin levels. Thus, intermittent use of Car-200 along with Nat Sulph-30 yielded additional benefit against genotoxicity, cytotoxicity, hepatotoxicity and oxidative stress induced by the carcinogens during hepatocarcinogenesis.

Molecular targeting for treatment of advanced hepatocellular carcinoma / 대한간학회지

Hepatocellular carcinoma (HCC) is a major global health problem, which has a grave morbidity and mortality. Over the past few decades, no effective systemic therapeutic modalities have been established for patients with the unresectable HCC in advanced stage. Sorafenib is a small molecule that blocks cancer cell proliferation by targeting the intracellular signaling pathway at the level of Raf-1 and B-Raf serine-threonine kinases, and exerts an anti-angiogenic effect by targeting the vascular endothelial growth factor receptor-1, 2 and 3, and platelet-derived growth factor receptor-beta tyrosine kinases. Recently, two clinical successful applications, SHARP and Asia-Pacific trial, of multikinase inhibitor sorafenib represent a significant advance in the treatment of advanced HCC patients without a curative chance. However, because the results of clinical trials show diverse responses in a subset of HCC patients, a molecular classification of HCC through the excavation of specific biomarkers related to its biological behavior is necessary for sorting HCC patients to each group with a biological homogeneity, ultimately leading to the most suitable individualization of molecular targeted therapy in HCC.

Atividade quimiopreventiva da tributirina quando administrada isoladamente ou em associação com vitamina A na etapa de promoção da hepatocarcinogênese em ratos / Chemopreventive activity of tributyrin when administrated isolated or in association with vitamin A in promotion phase of hepatocarcinogenesis to rats

No presente estudo avaliou-se a atividade quimiopreventiva da vitamina A (VA), tributirina (TB) e/ou 5-azacitidina (5-AzC) quando administradas isoladamente ou em associação na etapa de promoção do modelo de hepatocarcinogênese. Para tanto, ratos Wistar foram submetidos ao modelo do "hepatócito resistente" (HR), que consistiu na aplicação intraperitoneal de uma dose do agente iniciante dietilnitrosamina (DEN, 20 mg/100 g de p.c.), seguida, 2 semanas após, da aplicação de 4 doses consecutivas de 2- acetilaminofluoreno (2-AAF; 2 mg/100 g de p.c.) e de hepatectomia parcial (HP) a 70 ’POR CENTO’, acrescida de 1 dose de AAF (0,5 mg/100 g de p.c.) 4 dias após a cirurgia. Logo após a aplicação do modelo, os animais receberam VA (1 mg/ 100 g de p.c.) em dias alternados, TB (200 mg/ 100 g de p.c.) todos os dias e /ou 5-AzC (0,25 mg/ 100g de p.c.) duas vezes por semana ou, ainda, óleo de milho, maltodextrina e solução salina (grupo controle) durante cinco semanas. De acordo com a análise macroscópica do fígado, e em comparação ao grupo controle, verificou-se que o grupo tratado com TB isoladamente e o grupo tratado com VA e TB em associação apresentaram menor (p < 0,05) incidência e menor (p < 0,05) número médio de nódulos hepáticos. Em relação à análise morfométrica das lesões pré-neoplásicas (LPN) hepáticas positivas para a enzima glutationa S-transferase forma placentária (GST-P), observou-se que em comparação ao grupo controle os grupos experimentais que receberam 5-AzC apresentaram maior (p < 0,05) número de LPN hepáticas totais (persistentes + remodelação), LPN persistentes e LPN em remodelação, indicando que a 5-AzC teve ação potencializadora da iniciação nesse modelo de hepatocarcinogênese. Além disso, quando comparado ao grupo controle, observou-se que o grupo que recebeu TB isoladamente e o grupo que recebeu TB e nos tamanhos das LPN persistentes. Em comparação aos animais do grupo controle, não...

Chemopreventive activity of vitamin A (VA), tributyrin (TB) and/or 5-azacitidine (5-AzC) when administrated isolated or in association, was evaluated during the promotion phase of the "Resistant Hepatocyte"(RH) model of hepatocarcinogenisis. Rats received one dose of diethylnitrosamine (DEN; 20 mg/100g body weigh [b.w.]) for initiation and two weeks later, the animals received five doses of 2-acetylaminofluorene (AAF, 2 mg/100g b.w.), 4 consecutive doses before partial (2/3) hepatectomy and the remaining one four days after surgery. After 3 days, the animals received vitamin A (1mg/100 g.w.), TB (200 mg/ 100g de b.w.) and /or 5-AzC (0,25 mg/ 100 g de b. w.) or corn oil, maltrodextrin and saline (control goup) during five consecutive weeks. Incidence area of visible hepatocyte nodules/animal were smaller (p < 0,05) in goups that received TB and VA +TB than control group. Number of total (persistant + remodeling), persistent and remodeling hepatic placental glutathione S-transferase (GST-P) positive preneoplastic lesions (PNL) were higher (p < 0,05) in groups that received 5-AzC, indicating that 5-AzC potentiates initiation induced by carcinogens in rat liver. Moreover, the groups that received TB isolated or in association with VA presented mean area of persistent hepatic GST-p positive PNL smaller (p < 0,09) than control group. The animals treated with 5-AzC showed higher (p < 0,05) growth index in PNL than control group. The treatment with VA and TB isolated or in association increased (p < 0,05) the number of apoptotic bodies in PNL when compared to control group. Compared to normal group lower (p < 0,05) hepatic concentrations of retinyl palmitate were observed in control group, indicating that VA metabolism is altered in initial phases of hepatocarcinogenesis. The experimental groups that received VA showed higher concentrations of retinol and retinyl palmitate than control group. Moreover, the animals that received TB showed higher concentration...

Ácido fólico: efeitos paradoxais na promoção da hepatocarcinogênese em ratos / Folic acid: paradoxical effects during promotion of hepatocarcinogenesis in rats

A suplementação com ácido fólico (AF) apresenta efeitos quimiopreventivos, porém, pode aumentar o risco de desenvolvimento e acelerar a progressão do câncer se ocorrer em doses elevadas ou após a ocorrência de lesões pré-neoplásicas (LPN). O AF é essencial na síntese de novo de purinas e timidalato e consequentemente na síntese, replicação e reparo do DNA, proliferação celular e apoptose. Assim, a deficiência pode implicar em danos ao DNA e erros na sua replicação e reparo, processos importantes na carcinogênese, onde as células apresentam taxas de replicação e divisão aceleradas, e é possível que a suplementação module estes processos. Além disso, como AF ocupa uma posição de destaque no metabolismo dos grupamentos metila pode exercer efeitos sobre a hipometilação global do DNA e o aumento da expressão de proto-oncogenes como o c-myc, fenômenos característicos da hepatocarcinogênese. Assim, objetivando-se avaliar os efeitos do AF na promoção da hepatocarcinogênese em ratos Wistar, desenvolveu-se o modelo do /"Hepatócito Resistente/" e administrou-se por entubação gástrica diariamente, durante 5 semanas, o AF (0,16; 0,32; ou 0,64 mg / 100 g de peso / dia) ou água (0,25 mL / 100 g de peso / dia). Então, avaliou-se as LPN hepáticas presentes visíveis à macroscopia e microscopia (GST-P), a proliferação celular (BrdU) e a apoptose (microscopia de fluorescência) no tecido hepático ao redor das LPN e nas LPN persistentes e em remodelação, a intensidade de danos ao DNA (/"Cometa/" alcalino), e o padrão de metilação global (Dot Blot) e a expressão do c-myc (RT-PCR) especificamente em LPN microdissecadas. Apesar de não ter alterado a incidência e multiplicidade das LPN, o tratamento com AF 0,32 mg / 100 g promoveu um aumento na porcentagem de lesões ≥ 1 mm e o com AF 0,64 mg / 100 g a diminuição na porcentagem dessa lesões com relação ao grupo água (p<0,05). De modo semelhante, observou-se na análise das LPN GST-P positivas que o AF 0,32 mg / 100 g promoveu aumento e...

Folic acid (FA) supplementation shows chemopreventive effects, however, it may increase the risk of development and accelerate cancer progression in case of high doses or after preneoplastic lesions (PNL) are established. FA is essential on de novo synthesis of purine and thymidalate and, consequently, on DNA synthesis, replication and repair, cell proliferation and apoptosis. Thus, its deficiency may cause DNA damage and replication and repair mistakes, important processes on carcinogenesis, where cells present high replication rates and accelerated division, and is possible that supplementation modulates these processes. Besides, as FA has a central role on methyl group metabolism, it may have effects on hepatocarcinogenesis peculiar events such as DNA global hypomethylation and on the increased expression of proto-oncogenes like c-myc. Objecting the evaluation of FA effects during hepatocarcinogenesis promotion in Wistar rats, the /"Resistant Hepatocyte/" model was developed and water (0.25 mL / 100 g BW / day) or FA (0.16; 0.32; or 0.64 mg / 100 g BW / day) were supplemented daily by gavage for 5 weeks. Then, hepatic PNL detected by macroscopy and microscopy (GST-P), cell proliferation (BrdU) and apoptosis (fluorescence microscopy) on surrounding tissue, persistent and remodeling PNL, DNA damage (alcaline Comet assay), DNA global methylation pattern (Dot Blot) and c-myc expression (RT-PCR) specifically in microdissected PNL were evaluated. Even though FA treatment was not able to change incidence and multiplicity of PNL, the treatment with 0.32 mg / 100 g of FA increased the percentage of lesions ≥ 1 mm whereas with 0.64 mg / 100 g of FA diminished the percentage of these lesions, compaired to the water group (p<0.05). Similarly, it could be observed in PNL positive GST-P analysis that FA 0.32 mg / 100 g enhanced and FA 0.64 mg / 100 g inhibited the carcinogenic process, although it was not possible to detect significant differences on number, size and…

Histopatología de cirrosis y hepatocarcinoma por Virus B / Cirrhosis and hepatocarcinoma in HBV

Cirrosis por virus B: Se puede detectar en biopsia el antígeno de superficie con tinción de orceína (hepatocitos esmerilados) y por inmunohistoquímica (antígeno de superficie y anticore). Estadios de evolución: a) temprano o incipiente: nódulos no definidos, mucha fibrosis y alteración arquitectural; b) establecido: nódulos rodeados completamente de bandas fibroticas. estadio histológico: a) activo: necrosis e inflamación importante, colestasis; b) Inactivo: poca necrosis o inflamación, hepatocarcinoma: (tipos): trabecular, células claras, adenoide, sólido, escirroso. La immunohistoquimica ayuda a definir el tipo usando los antígenos hepatocelulares, antígeno carcinoembriónico, citoqueratinas, cromogranina y sinaptofisina.

HBV cirrosis: The surface antigen can be detected by biopsy, orcein tinctions (emery hepatocytes), and immunohistochemistry (surface antigen and anticore). Evolution Stages: a) Early or incipient (non-defined nodes, a lot of fibrosis and architectural alteration); b) established (nods completely surrounded by fibrotic bands). Histological Stages: a) active (main necrosis and inflammation; and cholestasis); b) inactive (little necrosisor inflammation). Types of Hepatocarcinomas: trabecular-clear cells- adenoid-solid-scirrhous. The immunohistochemistry helps to define the type using the hepatocellular antigens, carcinoembryonic antigen, cytokeratin, chromogranins and synapto-physins.

Pre- and post-initiation modulating effects of green tea ingestion on rat hepatocarcinogenesis

The purpose of this study was to investigate the effects of green tea ingestion on hepatocarcinogenesis before and after its initiation. Male Sprague-Dawley rats were fed an AIN76A diet with or without green tea. Initiation was induced by a single dose (200 mg/kg) of diethylnitrosamine at week 4 and 0.02% (w/w) 2-acetylaminofluorene was supplied in the diets. The control group had free access to water for 13 weeks (CTR13). Tea infusion was provided from the beginning of the experiment for 13 weeks (PRE13) or from the post-initiation stage until week 13 (POST13). Three other groups (CTR24, PRE24 and POST24) were added to examine the longer-term effects (24 weeks) with the same experimental design. The percentage area of liver sections that were positive for hepatic placental glutathione S-transferase (GST-P), which was used as a marker of preneoplastic lesions, was smaller in PRE13 (20.2 +/- 5.0%, mean +/- SD) and POST13 (26.0 +/- 4.8%) than in CTR13 (33.2 +/- 5.8%, p<0.05). Over the longer period, the GST-P lesions were significantly smaller for both PRE24 and POST24 (21.6 +/- 8.5% and 22.2 +/- 4.0%, respectively) than for CTR24 (28.6 +/- 5.1%, p<0.05), but there was no significant difference between PRE24 and POST24. The liver content of thiobarbituric acid reactive substances was significantly lower in the tea groups than in the controls (p<0.05). However, no significant differences were observed among groups of GST activity. The results show that tea consumption exhibits a stronger short-term initiation-inhibiting ability in liver carcinogenesis, but over a longer period, the preventive effects of green tea ingestion do not differ in post- and pre-initiation.

Ginger Extract (Zingiber officinale Roscoe) Triggers Apoptosis in Hepatocarcinogenesis Induced Rats

Ginger extract has been reported previously by our group to exhibit anticancer and antioxidant effects by reducing tumour burden and lipid peroxidation respectively in he-patocarcinogenesis induced rats. The current study examined the expression of pro-apoptotic protein caspase-8 and anti-apoptotic protein Bcl-2 in hepatocarcinogenesis treated rats. Thirty normal male Wistar rats were divided into 5 groups based on the diet given: i) control (normal rat chow), ii) olive oil, iii) ginger extract (100mg/kg body weight), iv) choline deficient diet + ethionine, CDE (to induce liver cancer) and v) CDE+ginger extract. Rats were killed at week 8, and liver tissues were excised for immuno-histochemical study to identify pro-apoptotic and anti-apoptotic proteins, caspase-8 and Bcl-2. The observation on H&E staining confirmed the CDE diet induced liver can-cer as indicated by the presence of numerous oval cells. Identification of Bcl-2 expres-sion showed that 91.6% (11/12) of the samples from the CDE group revealed positive staining while treatment with ginger extract however inhibited the expression with only 8.4% (1/12) samples showing positive staining for Bcl-2. As for caspase-8 protein, 41.7% (5/12) of the samples from CDE group showed positive staining, which in-creased to 100% (12/12) with ginger extract treatment. Our findings suggest that gin-ger extract has an anticancer effect by inducing apoptosis in liver cancer cells via up-regulation of the expression of pro-apoptotic protein, caspase-8 and down-regulation of the expression of anti-apoptotic protein Bcl-2.

Effects of Vitamin E and Dehydroepiandrosterone on The Formation of Preneoplastic Lesions in Rat Hepatocellular Carcinogenesis

This study is designed to examine the effects of dietary supplementation with vitamin E and dehydroepiandrosterone (DHEA) on the formation of preneoplastic lesions in diethylnitrosamine (DEN) induced rat hepatocarcinogenesis. All Weaning male Sprague-Dawley rats were initiated by a single dose of DEN (200 mg/kg body weight), subjected to twothirds partial hepatectomy 3 weeks later and were sacrificed 8 weeks after DEN initiation. Two weeks after initiation, rats were fed Purina purified rodent diet 5053 (Ralston Purina Rat chow, USA) with 1.5% (15,000 IU/kg diet) vitamin E, 0.5% DHEA and both of those supplemented diet for 6 weeks. Placental glutathione S-transferase (GST-P) positive foci, the activities of catalase, total-glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST) and thiobarbituric acid reactive substances (TBARS) contents were decreased significantly by vitaimin E supplement. On the other hand GST-P positive foci number, Cu/Zn-superoxide dismutase (SOD) and glucose 6-phosphatase (G6Pase) activities weren't changed by vitamin E supplement. It might suggest that protective effect of vitamin E against hepatocarcinogens is not involved in the formation of the GST-P positive foci but related to the expansion of that. It seemed that vitamin E supplement helped endogenous defense system in carcinogenesis by decreasing TBARS contents, H2O2, organic peroxides. Therefore, vitamin E seemed to protect cell from free radical damage in carcinogenesis . By DHEA supplement liver weight and liver/body ratio were increased, the area and number of GST-P positive foci, the activities of catalase, GR, total GPx, GST and the TBA RS contents were decreased significantly. On the other hand Cu/Zn-SOD and G6Pase activities weren't changed by DHEA supplement. In hepatocarcinogenesis the activities of antioxidant enzymes weren't increased by DHEA supplement. DHEA did not increase the oxidative stress, while DHEA seems to have anticarcinogenic effect in rats hepatocarcinogenesis.