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Item Supernatant of bacterial fermented soybean induced apoptosis of human hepatocellular carcinoma Hep 3B cells via activation of caspase 8 and mitochondria(ELSEVIER, 2007-02-01) Su, C.-L.*, Wu, C.-J., Chen, F.-N., Wang, B.-J., Sheu, S.-R., and Won, S.-J.,SC-1, the aqueous phase of soybean fermentation products by bacteria (Bacillus subtilis and Bacillus brevis), significantly inhibited the growth and clonogenesity of human hepatocellular (Hep 3B), mouse hepatocellular (ML-1), and human colorectal (HCT 116 and HT-29) carcinoma cells. Cytotoxicity of SC-1 in Hep 3B cells was through the process of apoptosis characterizing by increase in cell population of sub-G1 phase, fragmentation of DNA, and change of nuclear morphology. Treatment of Hep 3B cells with SC-1 activated caspase 8 and caspase 3. Elevation of nuclear DNA fragmentation factor 40 (DFF40) and cleavage form of poly(ADP-ribose) polymerase (PARP) were also observed. SC-1 also activated intrinsic pathway via increase of pro-apoptotic (tBid, Bak and Bax) and decrease of anti-apoptotic (Bcl-2 and Bcl-xL) proteins on mitochondria, disruption of mitochondrial membrane potential, release of cytochrome c and Smac (second mitochondria-derived activator of caspase/direct IAP binding protein with low PI) from mitochondria, and activation of caspase 9. Inhibition on protein expression of Ku70 in cytosol and cyclooxygenase (COX)-2, but not COX-1, in whole cell lystes were revealed in SC-1-treated Hep 3B cells. These results suggest caspase 8, Ku70 and mitochondria are involved in the antitumor mechanism of SC-1 in Hep 3B cells.Item Free radical scavenging and apoptotic effects of Cordyceps sinensis fractionated by supercritical carbon dioxide(ELSEVIER, 2005-04-01) Wang, B.-J., Won, S.-J., Yu, Z.-R., and Su, C.-L.,Supercritical carbon dioxide (SC-CO2) was used as the elution solvent for fractioning ethanolic extract (E) of Cordyceps sinensis (CS), a traditional Chinese herbal remedy, into R, F1, F2, and F3 fractions. This extractive fractionation method is amenable to large scale and is nontoxic. These four fractions were characterized in terms of total polysaccharides and cordycepin concentrations, scavenging ability of free radicals, and anti-tumor activities. Experimental results demonstrated that fractionation altered the distributions of total polysaccharides and cordycepin in fractions. Fraction R was the most active fraction to scavenge free radicals and inhibit the proliferation of carcinoma cells, followed by the fraction F1 and the extract E. The effect of scavenging on 1,1-diphenyl-2-picryl hydrazyl (DPPH) of CS extract and fractions at 2 mg/ml was R (93%), F1 (75%), E (66%), F2 (47%), and F3 (27%). The IC50 (50% cell growth inhibitory concentration) of tumor cell proliferation and colony formation on human colorectal (HT-29 and HCT 116) and hepatocellular (Hep 3B and Hep G2) carcinoma cells by fraction R were around 2 μg/ml. Conversely, R did not affect the growth of normal dividing human peripheral blood mononuclear cells (PBMC) by exhibiting a large value of IC50 over 200 μg/ml. Accumulation of tumor cells at sub-G1 phase and the fragmentation of DNA, typical features of programmed cell death, were observed in a time and dose dependent manner. Scavenging of free radicals and anti-cancer activity (value of IC50) correlated closely with the quantities of polysaccharides (Spearman’s rho = 0.901 and −0.870, respectively). Taken together, our findings suggest that fraction R, obtained by SC-CO2 fluid extractive fractionation, showed strong scavenging ability and selectively inhibited the growth of colorectal and hepatocellular cancer cells by the process of apoptosis.Item Caspase-8 acts as key upstream executor of mitochondria during justicidin A-induced apoptosis in human hepatoma cells(FEBS, 2006-05-01) Su, C.-L., Huang, L. L.-H, Huang, L.-M., Lee, J.-C., Lin, C.-N., and Won, S.-J.,Justicia procumbens is a traditional Taiwanese herbal remedy used to treat fever, pain, and cancer. Justicidin A, isolated from Justicia procumbens, has been reported to suppress in vitro growth of several tumor cell lines as well as hepatoma cells. In this study, justicidin A activated caspase-8 to increase tBid, disrupted mitochondrial membrane potential (Δψm), and caused the release of cytochrome c and Smac/DIABLO in Hep 3B and Hep G2 cells. Justicidin A also reduced Bcl-xL and increased Bax and Bak in mitochondria. Caspase-8 inhibitor (Z-IETD) attenuated the justicidin A-induced disruption of Δψm. Growth of Hep 3B implanted in NOD-SCID mice was suppressed significantly by oral justicidin A (20mg/kg/day). These results indicate that justicidin A-induced apoptosis in these cells proceeds via caspase-8 and is followed by mitochondrial disruption.Item Involvement of apoptosis and autophagy in reducing mouse hepatoma ML-1 cells growth in inbred BALB/c mice by bacterial fermented soybeans products(Elsevier, 2011-01-01) Su, C.-L., Chen, F.-N., and Won, S.-J.,Followed by the results of our previousn vitroeport (Food Chem. Toxicol., 2007), the efficacy of the soybean fermentation products containing live bacteria (SCB) was demonstrated using a syngeneic animal model. Murine HBV-related hepatoma ML-1 cells, derived from inbred animals and tumorigenic in BALB/c mice, were implanted subcutaneously to the flank of BALB/c mice on day 0. Three days after implantation, SCB (1.0 or 1.3l/mouse/day) or vehicle (water) was orally administrated daily until day 60. The results indicate that SCB significantly reduced (P�<�0.05) the volumes and weights of tumors during the experimental periods. Examination using TUNEL staining on section of tumors revealed apoptotic phenomenon of nuclear DNA double-strand breaks in the groups of mice received SCB. Immunohistochemistry further revealed an autophagic LC3-II punctate pattern. Of note, SCB induced autophagy in the absence or presence of apoptosis, whereas, apoptosis was observed only in combination with autophagy.n vitrotudy using autophagy inhibitor indicated that the induction of autophagy promoted apoptosis. These data imply that the suppression in tumor volumes and tumor weights by oral administration of SCB was due to the induction of apoptotic and autophagic cell death, which suggests therapeutic potential of SCB on HBV-related HCC.