Avalia??o do efeito do ?cido g?lico no tratamento de c?lulas de hepatocarcinoma HEPG2

• Main Author: Lima, Kelly Goulart
• Other Authors: Oliveira, Jarbas Rodrigues de
• Format: Others
• Language: Portuguese
• Published: Pontif?cia Universidade Cat?lica do Rio Grande do Sul 2015
• Subjects: BIOLOGIA CELULAR; BIOLOGIA MOLECULAR; ?CIDO G?LICO; C?LULAS; CARCINOMA HEPATOCELULAR; NEOPLASIAS; CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
• Online Access: http://tede2.pucrs.br/tede2/handle/tede/5495

Made available in DSpace on 2015-04-14T14:51:33Z (GMT). No. of bitstreams: 1 459268.pdf: 1089012 bytes, checksum: 3e3bbefbe5c53d842c73d0dec81136ac (MD5) Previous issue date: 2014-02-18 === Hepatocellular Carcinoma is the most prevalent primary tiver tumor and is among the top ten cancers that affect the world population. lts development is related, in most cases, the existente of chronic tiver injury, such as occurs in cirrhosis. Current curative therapies are only surgical resection, transplantation and percutaneous ablation, however with the possibility of recurrence. In this context, the search for new therapies for the disease becomes an interesting field for research. The knowledge about the correlation between chronic inflammation and cancer has driven new researches with anti-inflammatory agents that have potential for the development of antitumor drugs. Gallic acid is a polyphenol found in many natural products and have shown anti-inflammatory activity, anti-tumor, antimutagenic and strong antioxidant action. The purpose of this study was to investigate the effect of gallic acid on cell proliferation and inflammatory parameters of tiver carcinoma cells (HepG2), as well as to investigated the mechanisms involved. Cell viability was evaluated through MTT colorimetric assay and Trypan blue exclusion. Results showed that the gallic acid decreased proliferation of HepG2 celis in a dose and time dependent manner, without causing necrosis (LDH assay). We observed significant induction of apoptosis by PE Annexin V and 7-AAD assay and no interferente with the cell cycle using the FITC BrdU Flow Kit. The leveis of inflammatory mediators were measured by Cytometric Bead Array Human Inflammation Assay and observed a significant reduction in the leveis of interleukin-8 (pro-inflammatory and related to angiogenesis, invasiveness and metastasis) and increased leveis of interleukin-10 (anti-inflammatory and related to the induction of programmed cell death) and interleukin-12 (antiangiogenic and antimetastatic). We also evaluated the leveis of TGF by ELISA (Enzyme-Linked lmmunosorbent Assay) and no significant differences. According to these results, we believe that gallic acid has a strong potential as an anti-tumor agent. === O Carcinoma Hepatocelular ? o tumor hep?tico prim?rio mais prevalente e est? entre as dez principais neoplasias que afetam a popula??o mundial. O seu desenvolvimento est? relacionado, na maioria das vezes, a uma les?o hep?tica cr?nica, como ocorre na cirrose. As terapias consideradas curativas atualmente s?o somente a ressec??o cir?rgica, o transplante e a abla??o percut?nea, ainda com possibilidade de recidiva. Nesse contexto, a busca por novas alternativas terap?uticas para a doen?a torna-se um campo de pesquisa em expans?o. O conhecimento sobre a correla??o entre a inflama??o cr?nica e c?ncer tem impulsionado novas pesquisas com agentes anti-inflamat?rios que apresentem potencial para o desenvolvimento de drogas antitumorais. O ?cido g?lico ? um polifenol encontrado em v?rios produtos naturais, o qual tem apresentado atividade anti-inflamat?ria, antitumoral, antimutag?nica e forte a??o antioxidante. O objetivo desse estudo foi avaliar o efeito do ?cido g?lico sobre a prolifera??o celular e par?metros inflamat?rios das c?lulas de carcinoma hep?tico (HepG2), assim como investigar os mecanismos envolvidos. A viabilidade celular foi avaliada atrav?s do ensaio colorim?trico MTT e contagem celular por exclus?o com Azul de Trypan. Os resultados mostraram que o ?cido g?lico reduziu a prolifera??o celular de maneira dose e tempo dependente, sem causar necrose (ensaio LDH). Observamos indu??o significante da apoptose atrav?s do ensaio PE Annexin V and 7-AAD e nenhuma interfer?ncia no ciclo celular utilizando o kit FITC BrdU Flow. Os n?veis de mediadores inflamat?rios foram medidos utilizando o kit Cytometric Bead Array Human Inflammation. Observamos redu??o significante nos n?veis da interleucina 8 (pr?-inflamat?ria e relacionada ? angiog?nese, invasividade e met?stases), aumento dos n?veis de interleucina 10 (anti-inflamat?ria e relacionada ? morte celular programada) e aumento dos n?veis de interleucina 12 (antiangiog?nica e antimetast?tica). N?s tamb?m avaliamos os n?veis de TGF(3 por ELISA (Enzyme?Linked lmmunosorbent Assay) e n?o observamos diferen?as significativas. A partir desses resultados, acreditamos que o ?cido g?lico tem forte potencial como agente antitumoral.