Quantitative Proteomics Reveals That the Inhibition of Na(+)/K(+)-ATPase Activity Affects S-Phase Progression Leading to a Chromosome Segregation Disorder by Attenuating the Aurora A Function in Hepatocellular Carcinoma Cells.
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.
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Zhongwei
Z
Xu
Department of Gastroenterology and Hepatology, The Third Central Hospital of Tianjin , Tianjin 300170, China.
Fengmei
F
Wang
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Fengxu
F
Fan
Affiliated Hospital of Chinese People's Armed Police Force , Tianjin 300162, China.
Yanjun
Y
Gu
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Nana
N
Shan
Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Xiangyan
X
Meng
Affiliated Hospital of Chinese People's Armed Police Force , Tianjin 300162, China.
Shixiang
S
Cheng
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Yingfu
Y
Liu
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Chengyan
C
Wang
Central Laboratory, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.
Yueying
Y
Song
Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard , No. 1 Huizhi Huan Road, DongLi District, Tianjin 300309, China.
Ruicheng
R
Xu
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D064096:Aurora Kinase A / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D016203:CDC2 Protein Kinase; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins; D002853:Chromatography, Liquid; D020090:Chromosome Segregation / Q000187:drug effects; D056741:Cyclin D / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects* / Q000235:genetics; D015398:Signal Transduction; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D016159:Tumor Suppressor Protein p53 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
D000970:Antineoplastic Agents; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D056741:Cyclin D; D004791:Enzyme Inhibitors; D009687:Nuclear Proteins; C490381:SMC2 protein, human; D016159:Tumor Suppressor Protein p53; D010042:Ouabain; C576917:AURKA protein, human; D064096:Aurora Kinase A; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases; D000254:Sodium-Potassium-Exchanging ATPase
AURKA; Na+/K+-ATPase; SILAC; hepatocellular carcinoma; ouabain; proteomics