碩士 / 國立屏東科技大學 / 食品科學系所 / 97 / Accordingly, nutraceutical will continuously leading the brand and position a critical issue of the global food market, while functional food derived from seafood just explode the highlight and exhibit a great potential at this special field. Synechococcus sp., a member of cyanobacteria family, containing phycocyanin (CPC) is one of the major light harvesting pigment-protein which exhibiting diverse antioxidant, anti-inflammatory and anticancerous properties. The main purpose of the current study is assessment of antitumor activity of CPC from Synechococcus sp. U2. The measurement of dose-dependent, and time course effect on antitumor activity of phycocyanin can be depicted by MTT assay, microscopic observation and cell apoptosis assay, respectively. The results showed that CPC was extracted with lysozyme and by freezing-thawing cycles, and purified by hydrophobic interaction chromatography with butyl sepharose column and anion chromatography using DEAE sepharose resin. The recovery efficiency of CPC from crude extract was above 56%. Three milligram pure CPC was obtained from 1 g of dried cell mass and the purity ratio (A620/A280) was 4.43.
The molecular mass of purified CPC was 103 kDa and has two subunits with apparent molecular mass of 20.4 and 18.5 kDa respectively by electrophoresis technology. In cell, the results indicated that 74% decrease in the proliferation of Hep G2 cells when treated with 500 µg/mL CPC up to 72 hrs and the IC50 was 299.1 µg/mL. By reverse phase microscope, cell morphology changed in shrinkaged and cluster of round cells were observed. The Hep G2 cell treated with CPC analyzed by Flow Cytometry suggested that in cell cycle, cells treated with 500 µg/mL CPC for 72h showed 20.16±0.57% in sub-G1. In annexin V-PI assay, the data suggested that 5.39±0.3% to 12.62±0.69% early apoptosis cells can be found when CPC were employed for 72 hours. Furthermore, the results demonstrated that the caspase 3 activity was enhanced by 6-fold from 2.99±0.58% to 20.04±0.39% when treated with the subject. Conclusively, the present study illustrated that CPC followed apoptosis pathway on Hep G2 cell lines.
| Identifer | oai:union.ndltd.org:TW/097NPUS5253014 |
| Creators | Kai-Tse Cheng, 鄭凱澤 |
| Contributors | Ean-Tun Liaw, 廖遠東 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 113 |
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