ABSTRACT
Many studies have shown the Na(+)/K(+)-ATPase (NKA) might be a potential target for anticancer therapy. Cardiac glycosides (CGs), as a family of naturally compounds, inhibited the NKA activity. The present study investigates the antitumor effect of ouabain and elucidates the pharmacological mechanisms of CG activity in liver cancer HepG2 cell using SILAC coupled to LC-MS/MS method. Bioinformatics analysis of 330 proteins that were changed in cells under treatment with 0.5 µmol/L ouabain showed that the biological processes are associated with an acute inflammatory response, cell cycle, oxidation reduction, chromosome segregation, and DNA metabolism. We confirmed that ouabain induced chromosome segregation disorder and S-cell cycle block by decreasing the expression of AURKA, SMC2, Cyclin D, and p-CDK1 as well as increasing the expression of p53. We found that the overexpression or inhibition of AURKA significantly reduced or enhanced the ouabain-mediated the anticancer effects. Our findings suggest that AURKA is involved in the anticancer mechanisms of ouabain in HepG2 cells.
Subject(s)
Antineoplastic Agents/pharmacology , Aurora Kinase A/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , Gene Expression Regulation, Neoplastic , Ouabain/pharmacology , S Phase/drug effects , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Animals , Aurora Kinase A/genetics , Aurora Kinase A/metabolism , CDC2 Protein Kinase , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/genetics , Carrier Proteins/metabolism , Cell Cycle Proteins , Chromatography, Liquid , Chromosome Segregation/drug effects , Cyclin D/antagonists & inhibitors , Cyclin D/genetics , Cyclin D/metabolism , Cyclin-Dependent Kinases/antagonists & inhibitors , Cyclin-Dependent Kinases/genetics , Cyclin-Dependent Kinases/metabolism , Female , Gene Regulatory Networks/drug effects , Hep G2 Cells , Humans , Liver Neoplasms/drug therapy , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Mice , Mice, Nude , Nuclear Proteins/antagonists & inhibitors , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , S Phase/genetics , Signal Transduction , Sodium-Potassium-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/metabolism , Tandem Mass Spectrometry , Tumor Suppressor Protein p53/agonists , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE: Quantitative determination was made of ethyl-p-hydroxybenzoate in Dracaena cochinchinensis extracted with two technologies. METHOD: The sample was resolved with methanol and isolated by TLC, purged with methanol. Tge sample solution was chroma to graphed on a C18 column with acetonitrile-1% acetic acid (31:69) as mobile phase, detecting at 257 nm and content was calculated with external standard method. RESULT: The standard curves of ethyl-p-hydroxybenzoate were linear in the range of 0.206-4.12 ng, r = 0.9998. The average recovery was 97.2% and RSD was 1.4%. CONCLUSION: The content of ethyl-p-hydroxybenzoate in D. cochinchinensis extracted with heating-tree technongy is higher than that with traditional technology.
Subject(s)
Dracaena/chemistry , Parabens/analysis , Plants, Medicinal/chemistry , Technology, Pharmaceutical/methodsABSTRACT
OBJECTIVE: To study the effects of rhubarb extract by the technology free of heating (RETFH) and the extracts from crude rhubarb and from prepared rhubarb on intestine movement and analgesia in mice, and to compare the effects each other. METHODS: Intestine movement was studied by determining the distance in intestine after administrating the extracts from different technology with ink as indicator. Analgesia effect was studied by hot plate test and torsion test induced by acetic acid for the extracts from different technology. RESULTS: Diarrhea effect of low dose (0.75 mg/g) RETFH was moderate, and equal to that of high dose (3.00 mg/g) extract from prepared rhubarb. Remarkable analgesia effect of low dose (0.50 mg/g) RETFH was equal to that of middle and high dose (2.00 mg/g, 4.00 mg/g) extracts from prepared rhubarb and high dose (4.00 mg/g) extract from crude rhubarb. CONCLUSION: 0.50 mg/g RETFH possessed obvious diarrhea and analgesia effects. To utilize the technology free of heating may decrease dosage of rhubarb and save medicinal resource.
