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Avaliação do potencial quimiopreventivo do óleo de Pequi (Caryocar brasiliense Camb.) na hepatocarcinogênese quimicamente induzida em camundongos / Evaluation of potential chemopreventive oil pequi (Caryocar brasieliense Camb) in chemically induced Hepatocarcinogenesis in miceSimone Morais Palmeira 12 November 2014 (has links)
A flora brasileira possui várias plantas com grande potencial quimiopreventivo contra processos neoplásicos, sendo uma delas o fruto do Pequi (Caryocar brasiliense Camb). Essa fruta da região central do Brasil contêm na sua polpa e principalmente no extrato do óleo da sua polpa, várias substâncias antioxidantes. Relatos científicos recentes indicam que as substâncias no Pequi estão relacionadas com a intensificação do sistema imunológico e a redução do risco de doenças degenerativas como o câncer. Portanto, o presente trabalho avaliou o potencial quimiopreventivo do óleo de Caryocar brasiliense contra lesões hepáticas pré-neoplásicas induzidas quimicamente pela dietilnitrosamina (DEN) em camundongos. O iniciador dietilnitrosamina (DEN) na concentração de 10ug/g foi injetado intraperitonialmente em camundongos de 14 dias de idade. Foram formados cinco grupos experimentais: C (controle sem nenhum tratamento); DEN (Dietilnitrosamina 10ug); OP400 (óleo de Pequi 400mg/kg); DEN+OP100 (DEN+100mg/kg de óleo de Pequi); e DEN+OP400 (DEN+400 mg/kg de óleo de Pequi). Estes três últimos grupos receberam o óleo a partir do 30º dia até o 189° dia de vida. Os parâmetros estereológicos densidade de volume (Vv) e volume total (VTot) das lesões pré-neoplásicas (LPN) foram avaliados juntamente com a expressão das citoqueratinas CK8/18. O óleo de C. brasiliense reduziu o volume total das lesões pré-neoplásicas em 51% no fígado dos camundongos e em 20% no número total de animais acometidos com estas lesões na dose de 400 mg/kg. Redução no número de perfis de focos de hepatócitos alterados (FHA) CK8/18 - positivos foram observados no grupo DEN+OP400. Estes efeitos foram atribuídos às substâncias antioxidantes como os carotenóides (com ou sem atividade pró-vitamina A) e vitamina C que possivelmente atuaram na fase de promoção inibindo a proliferação celular e também pela indução da remodelação dos FHA. Portanto, concluímos que o óleo de C. brasiliense possui efeito hepatoprotetor no desenvolvimento de lesões pré-neoplásicas em fígado de camundongos induzidos por DEN e com potencial para uso na prevenção do câncer hepático / The brazilian flora has several plants with large chemopreventive potential against neoplastic processes, being one of them the Pequi fruit (Caryocar brasiliense Camb). This fruit of the central region of Brazil contains at its pulp and especially in the oil extract of its pulp, various antioxidant substances. Recent scientific reports indicate that the substances in Pequi are related to the intensification of the immune system and the reduction of risk of degenerative diseases, such as cancer. Therefore, this study evaluated the chemopreventive potential of the Caryocar brasiliense oil against pre-neoplastic liver lesions chemically induced by diethylnitrosamine (DEN) in mice. The initiator diethylnitrosamine (DEN) at a concentration of 10ug/g was intraperitoneally injected into14 days of age mice. Five experimental groups were formed: C (control without any treatment); DEN (diethylnitrosamine 10ug); OP400 (oil Pequi 400 mg/kg); DEN+OP100 (DEN + 100 mg/kg of Pequi oil); and DEN+OP400 (DEN + 400 mg/kg of Pequi oil). These last three groups received oil from the 30th day to the 189th day of life. The stereological parameters volume density (Vv) and total volume (VTot) of pre-neoplastic lesions (PNL) were evaluated together with the expression of cytokeratins CK8/18. The oil of C. brasiliense reduced the total volume of pre-neoplastic lesions in the liver in 51% of mice and 20% in the total number of affected animals with these lesions at a dose of 400 mg / kg. Reduction in the number of foci of altered hepatocytes (FAH) CK8/18 profiles-positive were observed in DEN+OP400 group. These effects have been attributed to antioxidants substances such as carotenoids (with or without provitamin A activity) and Vitamin C which possibly acted on the promotion stage inhibitting cell proliferation and also by inducing remodeling of FHA. Therefore, we conclude that the oil of C. brasiliense has hepatoprotective effect on the development of pre-neoplastic lesions in the mice liver induced by DEN and with potential for use in the prevention of liver cancer
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Ausência de atividade quimiopreventiva por parte da vitamina A quando administrada a ratas na etapa de pós-iniciação da carcinogênese mamária / Lack of chemopreventive activity of vitamin A when administered to rats during the stage of post-initiation of mammary carcinogenesisLeticia Okamoto 11 February 2010 (has links)
Avaliou-se a eventual atividade quimiopreventiva por parte da vitamina A (VA) quando administrada a ratas Sprague-Dawley durante a etapa de pós-iniciação da carcinogênese mamária induzida pelo 7-12 dimetilbenz(a)antraceno (DMBA). Com exceção de 10 animais que constituíram um grupo controle à parte de ratas consideradas normais [grupo normal (N)], e que não foram submetidas a qualquer procedimento experimental durante todo o estudo, 40 ratas com 50 dias de idade receberam o agente carcinogênico DMBA, por meio de entubação gástrica na dose de 60 mg/kg/peso corpóreo. Após 2 semanas, as ratas iniciadas receberam durante 12 semanas consecutivas, por entubação gástrica, 0,25 mL/100 g de peso corpóreo de óleo de milho (grupo OM; controle, n=20) e 2,5 mg/100 g de peso corpóreo de vitamina A (grupo VA, n=20), sendo, então, eutanasiadas. Não houve diferenças (p>0,05) entre os grupos OM e VA quanto à latência de aparecimento da primeira neoplasia mamária, incidência, multiplicidade e peso médio das neoplasias mamárias, todas classificadas como malignas. Em comparação ao grupo OM, o grupo VA apresentou maior concentração (p<0,05) de retinol no tecido mamário neoplásico e hepático, além de maiores concentrações hepáticas (p<0,05) de palmitato de retinila. Não se detectou palmitato de retinila em neoplasias mamárias de ambos os grupos. Não houve diferença (p>0,05) entre neoplasias mamárias do grupo OM e o tecido mamário do grupo N quanto à metilação global do DNA. Em comparação a neoplasias mamárias do grupo OM, neoplasias do grupo VA apresentaram menor (p<0,05) metilação global do DNA. Conclui-se que a VA não apresentou atividades quimiopreventivas e seu metabolismo encontra-se alterado em neoplasias mamárias. / The potential chemopreventive activity of vitamin A (VA) was evaluated when administered to Sprague-Dawley rats during the stage of post-initiation of mammary carcinogenesis induced by 7-12 dimethylbenz(a)anthracene (DMBA). Except for 10 animals that constituted a separate control group of normal rats [normal group (N)], that were not subjected to any experimental procedure throughout the study, 40 rats with 50 days of age received the carcinogen DMBA by gavage at the dose of 60 mg/kg/body weight. After 2 weeks, the rats received for 12 consecutive weeks, by gavage, 0.25 mL/100 g body weight of corn oil (OM group, control, n = 20) or 2.5 mg/100 g body weight of vitamin A (VA group, n = 20), being then euthanized. There were no differences (p> 0.05) between the OM and VA groups regarding the latency of onset of first breast neoplasm, incidence, multiplicity and average weight of breast tumors, all classified as malignant. Compared to the OM group, the VA group had a higher concentration (p <0.05) of retinol in neoplastic breast tissue and liver as well as higher concentrations (p <0.05) of retinyl palmitate in the liver. Retinyl palmitate was not detected in breast tumors of both groups. There was no difference (p> 0.05) between breast tumours of OM group and mammary tissue of N group regarding global DNA methylation. Compared to breast tumors of OM group, breast tumors of VA group had lower (p <0.05) global DNA methylation. VA did not show chemopreventive activity when administered to rats during the stage of post-initiation of mammary carcinogenesis. Furthermore, the metabolism of VA is altered in breast tumors.
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Study of tumorigenesis by means of transgenic mouse models expressing RET/PTC3 rearrangement and E7 under control of bovine thyroglobulin promoter and CD1 mouse strain treated with acrylamideJin, Ling 23 June 2009 (has links)
Thyroid carcinomas are the most common endocrine tumors in humans. There are three major types of carcinomas of thyrocyte origin, including papillary, follicular, and anaplastic carcinomas. Papillary thyroid carcinoma (PTC) is the most common type of thyroid malignancy accounting 80% of thyroid cancer cases, and present several histologic variants, namely classical (45%), follicular (18%), solid, diffuse-sclerosing, cribriform, … .Specific genetic events represent early initiating and late triggering events. Several genetic lesions have been identified in various thyroid carcinomas and some of them are specifically associated to one type thyroid cancer. For instance, RET/PTC is the most common molecular event in the radiation-associated PTC in childhood. <p>In the first part of the work, we studied two transgenic mouse models: the Tg-RET/PTC3 (Tg-RP3) mouse and the Tg-E7 mouse. Both strains express the human origin transgene (RET/PTC3 rearrangement or E7) exclusively in the thyroid under the control of the bovine thyroglobulin promoter. <p>Our study of these two models showed:<p>In both E7 and RET/PTC3 mouse models, the thyroids exhibited hyperplasia with own 'oncogene-dependent' follicular cell characteristics. Small follicular cells with hyperchromatic nuclei with an increased nucleus/cytoplasm ratio were numerous in the E7 mice, and large cells with convex apical border, a decreased nucleus/cytoplasm ratio, a pale nucleus and dispersed chromatin were found in the RET/PTC3 mice. <p>At 6, 10 months and later on, E7 mice developed huge heterogeneous, normal functional thyroid goiter, with no tumor formation. <p>As in previous studies on transgenic RET/PTC3 mouse models, the generally encountered features such as solid tumours were present. We also observed conventional variant of human PTC at late age (since 11 month-old) with quite low incidence (4%). In addition to solid and conventional variant PTCs, 28% of mice developed a peculiar big size thyroid tumor pattern with “proliferative papillary cystic changes with spindle cells and remodelling” and macrophage infiltration in the cysts at as early as 2 month of age; this kind of tumor histologically resembles the rare human young age 'diffused sclerosing' variant PTC (DSVP), but disappeared after 6 month. The other peculiar tumor exhibits morphological similarity with another rare human FAP-associated (Familial Adenomatous colonic polyposis) cribriform PTC, which showed a mixed architecture of several histological patterns (solid, follicular, cribriform). At 6 months, 26% of mice presented the cribriform tumor pattern.<p>From the analyse of the proliferation index in the two models, we conclude that RET/PTC3 fusion protein over stimulates MAPK and Akt/PKB-signalling pathways, through Ras-Raf-Mek-Erk, Ras-PI3-K/Akt/PKB, particularly in the large cells which were strongly positive for three proliferation markers. E7 bypasses these two pathways, by directly binding to Rb1 protein and releasing the E2F transcription factor which induces cell proliferation. <p>So RET/PTC3 and E7 mice present several morphologic features which mimic human PTC tumors; RET/PTC3 could therefore be used as a partial model for human PTCs. <p>Further investigation of gene expression will allow the characterization of the molecular phenotype of the observed variants.<p>In the second part of the work, we attempted to generate by xenobiotic administration an in vivo model of thyroid carcinoma. Chronic exposure of CD1 mice to acrylamide in the drinking water during 6 and 8 months at doses of 3mg/kg per day similar to those causing thyroid tumorigenesis after 2 years in rats, did not induce any thyroid tumors whatever the level of thyroid stimulation. <p><p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished
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Evaluation of tumor suppressor gene p53, oncogene c-erbB-2 and matrix-metalloproteinase-9 as prognostic and predictive factors in breast carcinomaRahko, E. (Eeva) 15 May 2007 (has links)
Abstract
Breast carcinoma is the most common malignancy in females in western countries. Classical prognostic factors such as the size of a primary tumor and the presence or absence of axillary lymph node metastases, malignancy grade and hormone receptor status reflect the subsequent risk of disease recurrence after primary therapy and the need for adjuvant therapies. However, most breast carcinomas are detected in the early stage of the disease and the value of these classical prognostic factors is limited. There is also a great need to find new factors predicting the clinical efficacy of the anticancer drugs available. In this thesis tumor suppressor gene p53, oncogene c-erbB-2 and matrix metalloproteinase-9 were evaluated for their prognostic relevance in breast carcinoma patients treated in Oulu University Hospital, and matrix metalloproteinase-9 was also analyzed in women with premalignant lesions in the breast tissue in order to examine its role in breast carcinogenesis. Histological analyses were carried out from formalin-fixed, paraffin-embedded primary tumor specimens and p53, c-erbB-2 and matrix metalloproteinase-9 (MMP-9) statuses were systematically analyzed by immunohistochemistry.
P53 expression correlated with disease-free survival and overall survival in patients with early-stage breast carcinoma, regardless of adjuvant antiestrogen therapy. The co-expression of p53 and c-erbB-2 characterizes a tumor type with a clinically aggressive course of breast carcinoma. The clinical efficacy of anthracyline-based chemotherapy in metastatic carcinoma might be limited in patients with p53 expression in a primary tumor. When postmenopausal patients with lymph node metastases and receiving adjuvant antiestrogen therapy were examined, MMP-9 expression indicated a slightly greater risk of breast carcinoma recurrence in patients with estrogen receptor negative tumors. Hyperplastic breast tissue and invasive breast carcinoma lesions expressed some MMP-9 immunopositivity. However, the strongest positivity was seen in ductal carcinoma in situ samples, suggesting that MMP-9 participates in breast carcinogenesis in the preinvasive phase.
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Collagen XIII in cardiovascular development and tumorigenesisTahkola, J. (Jenni) 25 November 2008 (has links)
Abstract
Collagen XIII is a type II transmembrane protein, which has a short intracellular domain and a large, mainly collagenous ectodomain. It is located at many cell-matrix junctions and in focal adhesions in cultured cells and it has a function in cell adhesive processes.
Overexpression of collagen XIII molecules with an 83 amino acid deletion in part of the ectodomain leads to fetal lethality in Col13a1del transgenic mice. Doppler ultrasonography was performed at 12.5 days of gestation on fetuses resulting from heterozygous matings and matings between heterozygous and wild-type mice. Some fetuses had atrioventricular valve regurgitation (AVVR) and all of them were transgene positive. In addition, fetuses had pathological changes in functional parameters. Histological analysis showed the trabeculation of the ventricles to be reduced and the myocardium to be thinner in the fetuses with AVVR. Based on in situ hybridization (ISH), collagen XIII mRNA are normal constituents of these structures. Overexpression of mutant collagen XIII results in mid-gestation cardiac dysfunction in fetuses, and these disturbances in cardiac function may lead to death in utero. The heterozygous mice that were initially of normal appearance had an increased susceptibility to develop B cell lymphomas, which originated in the mesenteric lymph node. Collagen XIII protein was not detected in normal lymph nodes or in the lymphomas. The incidence of lymphomas was higher in conventional conditions than in a specific pathogen-free facility. In addition, the expression of collagen XIII was localized in the intestine and the basement membrane was highly abnormal. These findings suggest that collagen XIII is a critical determinant of lymphanogenesis.
Using ISH, antibody staining and RT-PCR techniques collagen XIII expression was analyzed during carcinogenesis in mice and in man. Collagen XIII expression increased during carcinogenesis in mice and in man. In the malignant process collagen XIII mRNA localized in the basal epithelium and in the invasive cells. According to antibody staining malignant invasive cells were positive. Results may reflect the disturbed adhesion of epithelial cells and ECM and that may affect the behaviour of the malignant cells, suggesting that collagen XIII has a significant role in the initiation of the invasion.
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Carbonic anhydrase in normal and neoplastic gastrointestinal tissues:with special emphasis on isoenzymes I, II, IX, XII, and XIVKivelä, A. (Antti) 13 June 2003 (has links)
Abstract
The carbonic anhydrases (CAs) catalyse the hydration of CO2 to bicarbonate at physiological pH. This chemical interconversion is crucial since HCO3- is the substrate for several biosynthetic reactions. Carbonic anhydrases are involved in many physiological processes connected with respiration and transport of CO2/bicarbonate between metabolising tissues and the lungs, pH homeostasis and electrolyte secretion in a variety of tissues/organs. The present work was undertaken to study the distribution and expression of CA isoenzymes in the normal and neoplastic gastrointestinal tissues.
The expression of CA I, II, IX and XII in the human intestine and colorectal tumours was investigated by immunohistochemistry and western blotting. In the present study, immunohistochemical methods were also used to examine the location of CA IX and XII in the human pancreas and pancreatic tumours. The expression of CA XIV in the murine liver and intestine was studied using immunostaining and northern blotting.
The present results suggest that transmembrane CA XII is absent from the small intestine, but is expressed in all segments of the normal large intestine. The positive signal for CA XII was confined to the basolateral plasma membranes of the epithelial cells of the surface epithelial cuff. In tumours, the signal for CA XII became stronger in the deep part of the lesion. The intensity of the immunostaining for CA I and II was clearly found to decrease in benign lesions and became very weak in malignant colorectal tumours. The reciprocal pattern of expression observed for membrane-associated (CA IX and XII) and cytoplasmic (CA I and II) isoenzymes in intestinal samples suggests that CA IX and XII may be functionally involved in tumour progression to malignancy and/or in invasion. CA I and II, which are thought to play important physiological roles in the normal colorectal mucosa, may not be required for growth of colorectal cancers and their expression consistently diminishes with progression to malignancy.
In the human pancreas CA IX and XII appeared to be sporadically expressed in the basolateral plasma membrane of the normal acinar and ductal epithelium. The increased expression of CA IX in hyperplastic ductal epithelium may contribute to the pancreatic tumourigenesis.
CA XIV was expressed in the hepatocyte plasma membrane and its localization on both apical and basolateral membrane domains suggests an important role for this isoenzyme in the regulation of ion and pH homeostasis in the liver.
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Etude du rôle directe de l'expression des protéines du virus de l'hépatite C sur la voie de signalisation intra-cellualire PI3K-Akt et de son implication dans le développement du carcinome hépato-cellulaire / Direct impact of the proteins expression of HCV on PI3K-Akt signaling pathway and involvement in the development of hepatocellular carcinomaImache, Mohamed 12 January 2016 (has links)
Le but du projet est l’étude des régulateurs de la voie de signalisation intracellulaire de la voie Pi(3)K-Akt au travers de l’analyse du suppresseur de tumeur PTEN (Phosphatase and tenson homolog) et de la sérine/thréonine kinase mTOR (Mamalian Target of Rapamycin). Cette étude à plusieurs objectifs :1. Modulation de la voie Akt par HCV sur des foies humains, foies de souris FL-N/35 au niveau basal dans un premier temps et lors d’un boost de la voie in vitro sur des cultures primaires d’hépatocytes murins.2. Etude de l’expression ainsi que des modifications post-traductionnelles des modulateurs de la voie PI(3)K dans un modèle murin exprimant (FL-N/35) ou non l’ORF complète du virus de l’hépatite C (VHC).3. Confirmer nos précédentes données avec l’invalidation de PTEN dans un modèle de souris KO pour PTEN.4. Etendre ses données au niveau moléculaire dans l’optique d’une étude mécanistique grâce à l’analyse in vivo, ex vivo et in vitro d’un modèle murin KO pour PTEN.5. Compléter cette étude par l’analyse des déterminants viraux impliqués dans la dérégulation de la signalisation intracellulaire grâce à l’étude de souris NS5A.6. Examiner l’impact de la dérégulation de la voie PI(3)K-Akt dans le développement des Carcinomes hépatocellulaires (CHCs) induit par le VHC. / The goal of myt thesis is to study regulators of intracellular signaling pathway of Pi route (3) K-Akt through the analysis of tumor suppressor PTEN (Phosphatase and tenson homolog) and serine / threonine kinase mTOR (Target of Rapamycin Mamalian). This study has several objectives:1. Modulation of the Akt pathway by HCV in human liver, mouse livers FL-N / 35 to the basal level in a first time and at a track boost in vitro on primary cultures of mouse hepatocytes.2. Study of the expression and post-translational modifications of modulators of PI path (3) K in a murine model expressing (FL-N / 35) or not the complete ORF of hepatitis C ( HCV).3. Confirming our previous data with the invalidation of PTEN in a knockout mouse model for PTEN.4. Extending its data at the molecular level with a view to a mechanistic study through analysis in vivo, ex vivo and in vitro of a knockout mouse model for PTEN.5. Complete this study by analyzing the viral determinants involved in the dysregulation of intracellular signaling through the NS5A mouse study.6. Examine the impact of deregulation of IP route (3) K-Akt in the development of hepatocellular carcinoma (CHCs) induced by HCV.
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Association entre le mutant p.R249S de p53 et la protéine HBx du virus de l’hépatite B dans les carcinomes hépatocellulaires / Association between the mutant p.R249S of p53 and the HBx protein of hepatitis B virus in hepatocellular carcinomaGouas, Doriane 13 December 2011 (has links)
La mutation R249S (mutant p.R249S) du gène TP53, caractéristique de l'exposition à l'aflatoxine B1 (AFB1), est la plus fréquente dans les carcinomes hépatocellulaires (CHC) et est dans la plupart des cas associée à une infection chronique par le virus de l'hépatite B (VHB). En effet, il existe une synergie entre ces deux facteurs de risque, augmentant ainsi le risque de développer un CHC. Dans une première partie, nous avons étudié les mécanismes moléculaires de cette synergie dans différents modèles cellulaires puis dans une deuxième partie nous avons utilisé une approche épidémiologique pour étudier l'interaction entre la mutation R249S et le VHB. Nous avons tout d'abord montré que p.R249S avait perdu ses fonctions liées à p53wt. D'autre part, p.R249S était capable de former un complexe protéique avec l'oncoprotéine virale HBx dans les cellules de CHC PLC/PRF/5. Dans la deuxième partie de ce travail, nos résultats montrent que la mutation R249S est détectable dans l'ADN du sérum de sujets asymptomatiques de Gambie rurale (Afrique de l'Ouest). Notre travail met en évidence des variations temporelles quantitatives de la mutation R249S. Ces variations dépendent des niveaux d'exposition d'AFB1 mais également de la présence du VHB, suggérant une interaction entre l'AFB1 et le VHB. Enfin, dans une autre étude menée en Gambie et basée sur le recrutement de sujets ayant développés un CHC ou non (contrôles) nos résultats montrent que la mutation R249S est fortement associée au gène HBX complet dans les CHC. Cette association pourrait ainsi expliquer en partie l'effet synergique observé entre l'AFB1 et le VHB. A terme, une cible critique pourrait être identifiée pour des interventions préventives ou thérapeutiques précoces sur les CHC dans les régions de forte incidence / R249S mutation (mutant p.R249S) of TP53 gene, characteristic of the exposure to aflatoxin B1, is the most frequent TP53 mutation in hepatocellular carcinoma (HCC) and is highly associated with chronic hepatitis B virus infection (HBV). Indeed, a synergistic effect exists between these two main risk factors, thus increasing the risk to develop HCC. In a first part, we have studied the molecular mechanisms of this synergy in different cellular models and then, in a second part we have used an epidemiology-based approach to investigate the interaction between the R249S mutation and HBV. Firstly, we have shown that p.R249S has lost p53wt functions. Moreover, p.R249S formed a protein complex with the oncoprotein HBx from HBV in the HCC cell line PLC/PRF/5. In the second part, our results show that R249S mutation is detectable in plasma DNA of asymptomatic subjects from the rural Gambia (West Africa). Our work shows quantitative variations of R249S mutation that are dependent on the levels of exposition to AFB1 but also on the presence of HBV, suggesting an interaction between AFB1 and HBV. Finally, in another study performed in The Gambia and based on subjects with HCC or not (controls), our results show that R249S mutation is highly associated with HBX complete gene in HCC. Therefore this association could explain in part the synergistic effect observed between AFB1 and HBV. Eventually, a critical target may be identified for preventive or early therapeutic interventions on HCC of high-incidence regions
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Targeted delivery of embelin to cancer cellsEmjedi, Zaakiyah Z. January 2013 (has links)
>Magister Scientiae - MSc / Apoptosis or programmed cell death is vital to the development of organisms as they
maintain the balance between cell death and cell growth. Failure to activate apoptosis has
been implicated in carcinogenesis and often results from the over expression of anti–cancer
proteins such as the X–linked inhibitor of apoptosis protein (XIAP). XIAP is over expresses
in certain cancers and is a potent inhibitor of the initiator caspase 9 and effector caspases 3
and 7. The increased expression of XIAP in cancer cells result in the resistance to apoptosis.
The control of XIAP is therefore considered as a target for anti–cancer drug development.
Embelin or 2,5–dihydroxy–3–undecyl–1,4–benzoquinoine is a dihydroxyquinone compound
that was previously shown to inhibit XIAP. This drug was discovered by structure based
computational screening. The binding of embelin to XIAP displaces XIAP from caspases,
consequently eliminating the inhibitory effect of XIAP on apoptosis.
The objective of this study was to develop a gold nanoparticle that can be used for the
targeted delivery of embelin to cancer cells thereby enhancing pro–apoptotic effects of the
pro–apoptotic drug, ceramide. XIAP expression levels were investigated by Western blot
analysis in a panel of human cancer cell lines available in the laboratory to identify two cell
lines that can be used as low and high XIAP expression controls. Gold nanoparticles were
synthesized and conjugated with embelin and a cancer targeting peptide with the amino acid
sequence LTVSPWY. The biconjugated nanoparticles were used to co–treat MCF7 and
HepG2 cells with ceramide. Apoptosis was quantified using flow cytometry. The uptake of
gold nanoparticles was investigated using HR–TEM and ICP–OES. This study showed that
gold nanoparticles conjugated with the LTVSPWY peptide is specifically targeted to and
taken up by cancer cells. Gold nanoparticles conjugated with embelin promoted ceramide
induced apoptotic cell death of cancer cells. However, it was observed that gold nanoparticles
biconjugated with the LTVSPWY peptide and embelin failed to enhance the pro–apoptotic
effects of ceramide. iii
This study successfully demonstrated that gold nanoparticles conjugated with embelin could
be used to enhance the effects of anti–cancer drugs using ceramide as an example.
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Modulation of colon carcinogenesis by dietary ω-6/ω-3 fatty acid ratios : a chemopreventive strategy?Abrahams, Celeste H. January 2015 (has links)
Philosophiae Doctor - PhD / The aim of this study was to determine whether dietary fats constituting specific ω-6/ω-3 fatty acids (FA) ratio has chemopreventive modulating effects on the development of colon cancer. Western diets intake of saturated FA (SATS) and ω-6 polyunsaturated FA (PUFA) are very high relative to low ω-3 PUFA consumption. This high ω-6 and low ω-3 FA intake, resulting in a high ω-6/ω-3 FA ratio, appears to have a promoting effect on disease outcome, whilst increased ω-3 FA intake exhibiting anti-cancer effects. An animal cancer model was employed to evaluate the effects of dietary fat ratios on chemically induced carcinogenesis during cancer promotion. This was to determine whether the FA diets have a promoting or inhibitory effect on early neoplastic lesions by quantifying aberrant crypt foci (ACF) development and monitoring the crypt cells proliferative and apoptotic indices. The expressions of genes associated with changes in cells redox balance were also assessed. Common dietary fats were combined to produce the dietary fat ratios: sunflower oil (S), borage oil (B) and fish oil (F). Combinations of these oils generated the different ω-6/ω-3 FA ratios: SB (ω-6/ω-3: 38:1), SF (ω-6/ω-3: 13:1) and SBF (10:1). To represent the Western diet's high ω-6/ω-3 FA ratio profile, S (ω-6/ω-3: 501: 1) was used as a control, and canola oil and olive oil as additional reference. The dietary fats had no toxic effects on the liver and kidney based on serum clinical biochemical measurements. Diets containing borage oil (SB and SBF diets), canola and olive oil decreased (p<0.05) the crypt multiplicity of large (≥7 crypts/focus) ACF, exhibiting anti-cancer effects by decreasing (p<0.05) the proliferative activity of the rat colon crypts. Borage oil's protective effect resulted from the enhanced supply of C18:3ω-6 that has anti-inflammatory and anti-proliferative properties. The observed decrease (p<0.05) in apoptosis in the ACF was also facilitated by the up- and down-regulation of DNA repair and DNA replication associated genes, Xpa and Ercc2 by borage oil, respectively. Canola oil and olive had the largest inhibitory effect on suppressing crypt multiplicity by reducing (p<0.05) proliferation in the colon. Both oils effected the up-regulation (p<0.05) of the expression of several oxidative stress and anti-oxidant defence genes mediating the regulation of cell proliferation. The increased supply of C18:1ω-9 (canola and olive) and total polyphenolic content (olive) protected cells against oxidative stress induced apoptosis, which provided interesting interactive effects between FA and polyphenolic oil constituents that should be further elucidated. In contrast, the fish oil containing (SF diet) and the control sunflower (S diet) increased (p<0.05) the total ACF and colon crypt multiplicity (≥7 crypts/focus) when compared to the SB, SBF, olive oil and canola oil diets. An increased resistance to oxidative stress induced apoptosis appears to facilitate fish oil’s enhancing effect on crypt multiplicity despite the increased supply of LC ω-3 FA, which are prone to oxidation and leads to increased oxidative stress. This protective effect on crypt multiplicity and ACF development was mainly due to enhanced cellular antioxidant and DNA repair responses through the up-regulation (p<0.05) of Gpx4 and Nudt1, which favoured the increase (p<0.05) of crypt cells proliferation.The in vitro study demonstrated that oil ratio emulsions (S: ω-6/ω-3 = 249:1; SB: ω-6/ω-3 = 28:1; SF: ω-6/ω-3 = 12:1 and SBF: ω-6/ω-3 = 12:1) had differential effects on the survival indices of HT-29 and Caco-2 colon cancer cells. Contrary to the in vivo model, fish oil (SF and SBF emulsions) significantly (p<0.05) reduced the viability and proliferation of both cell lines, with the HT-29 cells showing greater sensitivity to the oil’s anti-proliferative effect. The HT-29 cells exposure to increased levels of C20:5ω-3 and C22:6ω-3 predisposes it to lipid peroxidation that increases the potential for cell removal via apoptosis. However, apoptotic effects were absent due to the HT-29 cells removal via necrosis as the cells energy status (ATP production) was significantly (p<0.05) depleted. Similar to the animal cancer model, borage oil (SB and SBF emulsions) had a reducing (p<0.05) effect on cell proliferation in both cell lines. However, as ATP was decreased (p<0.05), the S, SF and SBF emulsions resulted in an increased (p<0.05) apoptotic response in the Caco-2 cells in a dose dependent manner. This response resulted from the altered FA and lipid composition effected by the oil emulsions. Increased (p<0.05) incorporation of C20:5ω-3 and C22:6ω-3 in membrane phospholipid, phosphatidylethanolamine (PE), resulted in a significant decrease (p<0.05) in total SATS and MUFA content. A decrease (p<0.05) in membranes ω-6/ω-3 FA ratio was noted as well. This effect seems to selectively favour the induction of apoptosis by borage oil (SB and SBF). Similarly, an increase (p<0.05) in the PC/PE ratio by all oil emulsions, and a decrease (S and SB) and increase (SF and SBF) (p<0.05) in the chol/PL ratio appears to facilitate apoptosis too. A different threshold of the FA and lipid composition parameters elicits the inhibition of cell proliferation utilising lower oil emulsion concentrations. Therefore, the dietary supply of fats characterised by a defined low ω-6/ω-3 FA ratio can selectively modulate the growth indices of colon cancer. Specific oil ratio combinations by incorporating borage oil and fish oil hereby provide a selective strategy for chemoprevention in the colon, although underlying interactions and threshold effects of specific FA seems to prevail that should be further unravelled. / Medical Research Council (MRC) and Cancer Association of South Africa (CANSA)
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