title: Synergistic effect of simvastatin plus NS398 on inhibition of proliferation and survival in hepatocellular carcinoma cell line.
abstract: OBJECTIVE
  NS398, a selective cyclooxygenase-2 inhibitor, and simvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, both exert an anticancer effect on hepatocellular carcinoma cells, but the effect of co-administration of the two drugs remains unknown. We aimed to investigate the synergistic in vitro anticancer effect of co-administration of NS398 and simvastatin and its mechanism.
  
  
  METHODS
  The Hep3B and Huh-7 cell lines were cultured. Cells were treated with simvastatin, NS398, or a combination. 5-bromo-2'-deoxyuridine ELISA assay, flow cytometry, Western blot analyses, and immunofluorescence assay were performed.
  
  
  RESULTS
  In both cell lines, co-administration of simvastatin and NS398 resulted in a greater effect on proliferation and apoptosis. In Hep3B cells, co-administration of the two drugs resulted in a greater decrease in procaspase 3 and Bcl-2 and an increase in cleaved caspase 9 than that noted with monotherapy. In Huh-7 cells, co-administration of the two drugs resulted in a greater decrease in procaspase 3 and cyclin D1 and an increase in cleaved caspase 9. Expression of NF-κB and Akt were also decreased to a greater extent when the two drugs were co-administered in both cell lines. Immunofluorescence assay showed suppression of the nuclear localization of NF-κB by simvastatin or NS398. The effect was greater by co-administration.
  
  
  CONCLUSIONS
  The co-administration of NS398 and simvastatin produced greater antiproliferative and proapoptotic effects against Hep3B cells and Huh-7 cells. Inhibition of the NF-κB and Akt pathway and activation of caspase cascade, which are considered as the major mechanism of synergistic anticancer properties, were observed in both cell lines.
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, Korea University College of Medicine, Ansan, Korea.
authors.forename: Sun Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Ji Won
authors.initials: JW
authors.lastname: Hwang
authors.forename: Hyungshin
authors.initials: H
authors.lastname: Yim
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Yim
authors.forename: Sang Uk
authors.initials: SU
authors.lastname: Woo
authors.forename: Sang Jun
authors.initials: SJ
authors.lastname: Suh
authors.forename: Jong Jin
authors.initials: JJ
authors.lastname: Hyun
authors.forename: Sung Woo
authors.initials: SW
authors.lastname: Jung
authors.forename: Ja Seol
authors.initials: JS
authors.lastname: Koo
authors.forename: Ji Hoon
authors.initials: JH
authors.lastname: Kim
authors.forename: Yeon Seok
authors.initials: YS
authors.lastname: Seo
authors.forename: Jong Eun
authors.initials: JE
authors.lastname: Yeon
authors.forename: Sang Woo
authors.initials: SW
authors.lastname: Lee
authors.forename: Kwan Soo
authors.initials: KS
authors.lastname: Byun
authors.forename: Soon Ho
authors.initials: SH
authors.lastname: Um
mesh_terms: D000970:Antineoplastic Agents; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002471:Cell Transformation, Neoplastic / Q000187:drug effects*; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D004357:Drug Synergism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D016328:NF-kappa B / Q000378:metabolism; D009578:Nitrobenzenes / Q000494:pharmacology*; D051058:Oncogene Protein v-akt / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D019821:Simvastatin; D013449:Sulfonamides / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D052246:Cyclooxygenase 2 Inhibitors; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D016328:NF-kappa B; D009578:Nitrobenzenes; D019253:Proto-Oncogene Proteins c-bcl-2; D013449:Sulfonamides; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D019821:Simvastatin; D051058:Oncogene Protein v-akt; D053148:Caspase 3; D053453:Caspase 9
keywords: combination drug therapy; cyclooxygenase-2 inhibitor; hepatocellular carcinoma; simvastatin