ABSTRACT
The underground cane of Schizocapsa plantaginea (Hance) has long been used by Chinese ethnic minority as a constituent of anti-cancer formulae. Saponins are abundant secondary metabolic products located in the underground cane of this plant. The potential therapeutic effects of total saponins isolated from Schizocapsa plantaginea (Hance) (SSPH) on human hepatocellular carcinoma (HCC) were tested in vitro in human liver cancer cell lines, SMMC-7721 and Bel-7404. Apoptosis and cell cycle arrest were determined using flow cytometry, caspase activation was determined by ELISA, and PARP, cleaved PARP, mitogen-activated protein kinase (MAPK) expression and phosphorylation were measured using Western blotting analysis. In vivo anti-HCC effects of SSPH were verified in nude mouse xenograft model. SSPH exerted markedly inhibitory effect on HCC cell proliferation in time- and concentration-dependent manner. Moreover, SSPH significantly induced apoptosis through caspase-dependent signaling and arrested cell cycle at G/M phase. These anti-proliferation effects of SSPH were associated with up-regulated phosphorylation of extracellular signal-regulated kinase-1/2 (Erk1/2) and c-jun-NH2-kinase-1/2 (JNK1/2) and reduced phosphorylation of p38MAPK. Furthermore, inhibitors of ERK, UO126, and JNK, SP600125 inhibited the anti-proliferation effects by SSPH, suggesting that Erk and JNK were the effector molecules in SSPH induced anti-proliferative action. During in vivo experiments, SSPH was found to inhibit xenograft tumor growth in nude mice, with a similar mechanism in vitro. Our study confirmed that SSPH exerted antagonistic effects on human liver cancer cells both in vitro and in vivo. Molecular mechanisms underlying SSPH action might be closely associated with MAPK signaling pathways. These results indicated that SSPH has potential therapeutic effects on HCC.
Subject(s)
Animals , Humans , Mice , Antineoplastic Agents , Pharmacology , Toxicity , Apoptosis , Caspases , Genetics , Metabolism , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Cell Survival , Dioscoreaceae , Chemistry , Heterografts , Inhibitory Concentration 50 , Liver Neoplasms , Drug Therapy , Metabolism , Pathology , MAP Kinase Signaling System , Mice, Nude , Phosphorylation , Plant Tubers , Chemistry , Poly (ADP-Ribose) Polymerase-1 , Metabolism , Saponins , Pharmacology , ToxicityABSTRACT
The underground cane of Schizocapsa plantaginea (Hance) has long been used by Chinese ethnic minority as a constituent of anti-cancer formulae. Saponins are abundant secondary metabolic products located in the underground cane of this plant. The potential therapeutic effects of total saponins isolated from Schizocapsa plantaginea (Hance) (SSPH) on human hepatocellular carcinoma (HCC) were tested in vitro in human liver cancer cell lines, SMMC-7721 and Bel-7404. Apoptosis and cell cycle arrest were determined using flow cytometry, caspase activation was determined by ELISA, and PARP, cleaved PARP, mitogen-activated protein kinase (MAPK) expression and phosphorylation were measured using Western blotting analysis. In vivo anti-HCC effects of SSPH were verified in nude mouse xenograft model. SSPH exerted markedly inhibitory effect on HCC cell proliferation in time- and concentration-dependent manner. Moreover, SSPH significantly induced apoptosis through caspase-dependent signaling and arrested cell cycle at G/M phase. These anti-proliferation effects of SSPH were associated with up-regulated phosphorylation of extracellular signal-regulated kinase-1/2 (Erk1/2) and c-jun-NH2-kinase-1/2 (JNK1/2) and reduced phosphorylation of p38MAPK. Furthermore, inhibitors of ERK, UO126, and JNK, SP600125 inhibited the anti-proliferation effects by SSPH, suggesting that Erk and JNK were the effector molecules in SSPH induced anti-proliferative action. During in vivo experiments, SSPH was found to inhibit xenograft tumor growth in nude mice, with a similar mechanism in vitro. Our study confirmed that SSPH exerted antagonistic effects on human liver cancer cells both in vitro and in vivo. Molecular mechanisms underlying SSPH action might be closely associated with MAPK signaling pathways. These results indicated that SSPH has potential therapeutic effects on HCC.
Subject(s)
Animals , Humans , Mice , Antineoplastic Agents , Pharmacology , Toxicity , Apoptosis , Caspases , Genetics , Metabolism , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Cell Survival , Dioscoreaceae , Chemistry , Heterografts , Inhibitory Concentration 50 , Liver Neoplasms , Drug Therapy , Metabolism , Pathology , MAP Kinase Signaling System , Mice, Nude , Phosphorylation , Plant Tubers , Chemistry , Poly (ADP-Ribose) Polymerase-1 , Metabolism , Saponins , Pharmacology , ToxicityABSTRACT
<p><b>OBJECTIVE</b>To characterize properties of Rhizoma Dioscoreae polysaccharide-Fe (III) complex (RDPC) and analyze its composition.</p><p><b>METHOD</b>Physicochemical properties, such as character, solubility and stability of RDPC were studied. Surface structure of RDPC was studied by Infrared spectroscopic technique and transmission electron microscopy. The content of Fe (III) in RDPC was determined by o-phenanthroline spectrophotometry after being treated by ashing, wet digestion, or direct dissolution methods. Content of polysaccharide in RDPC was determined by sulphuric acid-phenol method.</p><p><b>RESULT</b>RDPC was brownish red powder. It could dissolve in water, its aqueous solution was very stable at the pH from 3.8 to 11.6. RDPC was a surface complex in which polysaccharide combined with beta-FeOOH as core. The content of Fe (III) in RDPC was 17.05%, 16.53%, 17.10% respectively after being treated by 3 different pretreatment methods. Content of polysaccharide in RDPC was 16.27%.</p><p><b>CONCLUSION</b>This is the first report of RDPC which was stable under physiological pH conditions and could be a new candidate for iron-supplementary.</p>