Gastric cancer is a high prevalent carcinoma and the leading cause of cancer-related mortality in Taiwan. Transforming growth factor-β is one of growth factors family, and involved in many biological processes leading to tumor formation, including cell proliferation, extracellular matrix secretion, cell adhesion, movement, differentiation and apoptosis. The TGF-β signaling its via two receptors complexes on the membrane, type I TGF-β receptor, TβRI and type II TGF-β receptor, TβRII, and activated downstream information by phosphorylation signaling mediator, protein Smad2/3, phosphorylated Smad2/3 was transferred to the nucleus to initiate transcription of downstream genes. Euphol is an euphane-type triterpene alcohol. It is isolated from the dichloromethane extract of Euphorbia, structurally similar to cholesterol which is a key component of lipid-raft microdomain on plasma membrane. It exhibits anti-viral and anti-inflammation activity, have higher cytotoxic in gastric cancer than normal cell, and will induce gastric cancer apoptosis pathway. However, the mechanisms and the potential of the anti-tumor properties of Euphol remain to be elucidated. The TGF-β receptor located in lipid-raft/caveolae microdomain stimulated by TGF-β will encounter lipid-raft/caveolae-mediated endocytosis, this endocytic pathway is a result of TGF-β receptors are transported to the lysosome for degradation. In contrast, TGF-β receptors which is located in non-lipid-raft region will be taken in the non-lipid-raft pathway. TGF-β receptors will transported into the early endosome, then going on signaling pathway. In this study, we found that TGF-β receptors were transferred from non-lipid raft to lipid-raft after Euphol treatment, this translocation increased degradation of TGF-β receptor and reduced the TGF-β signaling. Since the chemical structure of Euphol is similar to cholesterol. In our hypothesis, the cytotoxic of Euphol to gastric cancer may replace by cholesterol, then change the composed of protein and phospholipid, thereby influence the cell signaling of gastric cancer, then induce cancer cell death. Advisors/Committee Members: Yang, Jenq-Lin (chair), Chen, Chun-Lin (committee member), Huang, Yaw-Bin (chair).

A espécie Synadenium grantii Hook. f. é uma planta pertencente a família Euphorbiaceae, sendo conhecida popularmente como cega-olho, leitosinha, janaúba, cola-nota e outros. Seu látex tem sido usado tradicionalmente na região dos Campos Gerais na forma de uma garrafada, obtida da diluição deste em água (18 gotas em 1 litro). Suas aplicações medicinais são muito difundidas na cultura popular brasileira para a cura de vários tipos de câncer, além de outras enfermidades, como inflamações e úlcera. No entanto, não existem estudos científicos que comprovem esses efeitos além de pouco se saber sobre a constituição química da espécie. Assim, os objetivos deste trabalho foram realizar um estudo fitoquímico do látex de S. grantii; avaliar sua atividade antitumoral in vitro frente a linhagem celular de melanoma (B16F10); analisar as modificações na distribuição das células tumorais nas fases do ciclo celular e ainda avaliar a atividade antitumoral do látex in vivo bem como as alterações histológicas dos animais tratados com a forma popular de uso do látex da planta. A avaliação fitoquímica do látex foi realizada através de procedimentos cromatográficos (cromatografia em coluna e cromatografia em camada delgada) e espectroscópicos (RMN de 1H e 13C) da qual se obteve o isolamento e caracterização de um triterpeno e um esteroide identificados como eufol e citrostadienol respectivamente. A avaliação da citotoxicidade foi determinada pelo método de redução do MTT (brometo de [3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazólio) e pelo método de exclusão com Azul de Trypan. Para a realização dos ensaios in vitro, foram testadas concentrações do látex entre 1,7 μg/poço a 4,88 mg/poço e também as duas substâncias isoladas (Eufol e Citrostadienol) nas concentrações de 2,5 μg/mL - 250μg/mL. Para avaliação das modificações no ciclo celular, as células foram submetidas à análise por citometria de fluxo e a avaliação histológica foi realizada através da coloração hematoxilina-eosina. Os ensaios citotóxicos demonstraram que o látex de S. grantii foi capaz de diminuir o crescimento e a viabilidade das linhagens celulares em estudo apresentando efeito concentração e tempo dependentes nas duas metodologias avaliadas. Além disso, induziu parada do ciclo celular na fase S-G2/M. Das substâncias isoladas somente o citrostadienol apresentou uma reduzida atividade citotóxica in vitro frente a linhagem B16F10. Nos ensaios in vivo pode se observar uma diminuição em 40% no tamanho dos tumores dos camundongos tratados com a forma popular de uso do látex mas a avaliação histológica dos órgãos analisados (tumor, pulmão, fígado e linfonodos) foi semelhante no grupo tratado e não tratado.

The Synadenium grantii Hook f. species is a plant of the Euphorbiaceae family and is popularly known as cega-olho, leitosinha, janaúba, cola-nota among other names. Its latex has been used traditionally in the region of Campos Gerais as "garrafada", obtained from this dilution in water (18 drops into 1 liter). Its medicinal applications are widespread in…

Advisors/Committee Members: Giovani Marino Faveró, Flávio Luis Beltrame, Francinete Ramos Campos.

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The Synadenium grantii Hook f. species is a plant of the Euphorbiaceae family and is popularly known as cega-olho, leitosinha, janaúba, cola-nota among other names. Its latex has been used traditionally in the region of “Campos Gerais” as "garrafada", obtained from this dilution in water (18 drops into 1 liter). Its medicinal applications are widespread in Brazilian popular culture to cure various types of cancer, among other diseases, such as inflammation and ulcers. However, there are no scientific studies that prove these effects besides little is known about the chemical constitution of the species. The aims of this study was to conduct a phytochemical study of S. grantii latex; evaluate its antitumor activity in vitro against melanoma cell line (B16F10); analyze the changes in the distribution of tumor cells in cell cycle phases and also to evaluate the antitumor activity of latex in vivo and histological changes in animals treated with the popular way to use latex plant. The evaluation phytochemical latex was carried out by chromatographic procedures (column chromatography and thin layer chromatography) and spectroscopic procedures (1H NMR and 13C) from which was obtained the isolation and characterization of a triterpenoid and steroid identified as euphol respectively and citrostadienol. Evaluation of the cytotoxicity was determined by the reduction method of MTT ([3 - (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) and by exclusion with Trypan Blue. For the tests in vitro, látex concentrations of 0.012 μg/well to 48.8 mg/well were tested and also two isolated compounds (Euphol and Citrostadienol) at concentrations of 2.5 mg/mL - 250μg/mL. To assess changes in the cell cycle, the cells were analyzed by flow cytometry and histological evaluation was performed by hematoxylin-eosin. The cytotoxic assays demonstrated that the S. grantii latex was able to reduce the growth and viability of the cell lines showing concentration and time dependent effect in both methodologies evaluated. In addition, induced cell cycle arrest in S- G2/M phase. Isolated substances only citrostadienol showed a reduced cytotoxic activity in vitro against B16F10 lineage. In vivo assays demonstrated a reduction of 40% in the size of tumours in mice treated with the popular way to use latex but the histological evaluation of organs analyzed (tumor, lung, liver and lymph nodes) was similar in the treated and untreated animals.

A espécie Synadenium grantii Hook. f. é uma planta pertencente a família Euphorbiaceae, sendo conhecida popularmente como cega-olho, leitosinha, janaúba, cola-nota e outros. Seu látex tem sido usado tradicionalmente na região dos Campos Gerais na forma de uma “garrafada”, obtida da diluição deste em água (18 gotas em 1 litro). Suas aplicações medicinais são muito difundidas na cultura popular…

Advisors/Committee Members: Beltrame, Flávio Luis, Campos, Francinete Ramos, Faveró, Giovani Marino.