The Molecular Mechanism of Crude Extracts of Euchresta formosana Radix Induce Cell Cycle Arrest, Apoptosis and Inhibit Metastasis of Human Hepatocellular Carcinoma Cell Line （Hep3B）
| Autor: | Shu-Chun Hsu, 徐素琴 |
|---|---|
| Rok vydání: | 2007 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 95 Euchresta formosana (HAYATA) OHWI (Leguminosae) distributed in Java, Philippines and Taiwan occurring in shady and humid places. The roots of this plant have been used in detoxification, reduce inflammation, detumescence, analgesia, especially for the throat and snake wounds in Taiwan. Recently it can use in cancer therapy, but underlying mechanism is not well known. Our research to investigate the mechanism of anticancer activity of the 95% ethanol extracts from Euchresta formosana. Our data can be divided into two parts: the first part contains cell cycle arrest and cell apoptosis; the second part includes inhibition of metastasis. The data showed that the effects of 95% ethanol extracts of Euchresta formosana radix (EFR) on the cell cycle and apoptosis in human hepatocellular carcinoma (HCC) Hep3B cells were investigated. The results indicated that EFR decreased DNA synthesis and viable Hep3B cell numbers in a concentration-dependent manner. The EFR induced a p21- and p27-dependent cell cycle arrest in S-phase and apoptosis of the Hep3B cells. The induction of apoptosis by EFR treatment was also confirmed by DAPI staining. The EFR inhibited cyclin-dependent kinase (CDK) -1 and -2 expression and decreased cyclin B1 and E levels, resulting in S-phase arrest. The EFR induced reactive oxygen species (ROS) production followed by endoplasmic reticulum (ER) stress that was based on the increase of GADD153 and GRP78 which led to the release of Ca2+ in the Hep3B cells. The EFR promoted apoptosis was associated by increasing activation of caspase-3, -7, and -9 and enhanced poly(ADP-ribose) polymerase cleavage and increased expression of p21CIP1/WAF1, p27KIP1, Bax and Bad. Furthermore, the level of Bcl-xl decreased after EFR treatment. Alteration of these key anti- and pro-apoptotic proteins could contribute to the increase in p53-independent apoptosis that was observed in the Hep3B cells. The other phase, the effect of EFR on cell migration and invasion by the human hepatocellular carcinoma (HCC) cell line (Hep3B) was examined. Hep3B cells treated in vitro with EFR migrated and invaded less than cells treated with phosphate-buffered saline (PBS) as a control. EFR inhibited migration and invasion by down-regulating the production of RhoA and ROCK-1, FAK and matrix metalloproteinase-1, -2, -9 and -10 relative to PBS only. These results showed that EFR inhibits invasion and migration by liver cancer cells by down-regulating proteins associated with these processes, resulting in reduced metastasis. Thus, EFR should be considered as a possible therapeutic agent for inhibiting primary tumor growth and preventing metastasis. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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