Polyalthia longifolia is a lofty evergreen tree found in India and Sri Lanka. Today, P. longifolia var. pendula is in large-scale cultivation in south Taiwan as a landscape plant. A new compound, clerodane diterpenoid 16-hydroxycleroda-3,13-dien-15, 16 -olide¡]CD¡^, was isolated from the bark of Polyalthia longifolia var. pendula. The compound was shown with cell growth inhibitory property at first screen in KMU¡]Kaohsiung Medical University¡^. In our study, antiproliferative activity of CD on human colorectal adenocarcinoma cells¡]HT29 and HCT116¡^were tested and the inhibitory concertration of 50¢Mcell viability¡]IC50¡^is 5 £gg/ml¡]HCT116¡^ and 10 £gg/ml¡]HT29¡^determined by MTT(3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay. The caspase 3 and PARP cleavage experiment results indicated that CD induced apoptosis is dose dependent manner. We found that CD induced sub-G1 accumulation and reactive oxygen species(ROS) release by flow cytometry analysis. Pretreatment of N-Acetyl-L-cystine(NAC), an antioxidant agent, can reverse the anti-proliferation effect by CD. We found that CD induced loss of mitochondria membrane potentiol¡]£G£Zm¡^. We also analysis differentially expressed proteins by 2-D electrophoresis and find drug response proteins, e.g. HSP10, Profilin-1, Peroxindoxin-1. RT-PCR and western blot was performed to confirm the protein expression changes. It is interesting to reveal the role of these proteins in the colorectal cancinogenesis and anti-tumor drug response.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0715108-162559 |
| Date | 15 July 2008 |
| Creators | Chen, Chiu-Roung |
| Contributors | Hurng-Wern Huang, Yow-Ling Shiue, Chao-Neng Tseng |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0715108-162559 |
| Rights | off_campus_withheld, Copyright information available at source archive |
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