title: Silibinin induces apoptosis via calpain-dependent AIF nuclear translocation in U87MG human glioma cell death.
abstract: BACKGROUND
  Silibinin, a natural polyphenolic flavonoid, has been reported to induce cell death in various cancer cell types. However, the molecular mechanism is not clearly defined. Our previous study showed that silibinin induces glioma cell death and its effect was effectively prevented by calpain inhibitor. The present study was therefore undertaken to examine the role of calpain in the silibinin-induced glioma cell death.
  
  
  METHODS
  U87MG cells were grown on well tissue culture plates and cell viability was measured by MTT assay. ROS generation and △ψm were estimated using the fluorescence dyes. PKC activation and Bax expression were measured by Western blot analysis. AIF nuclear translocation was determined by Western blot and immunocytochemistry.
  
  
  RESULTS
  Silibinin induced activation of calpain, which was blocked by EGTA and the calpain inhibitor Z-Leu-Leu-CHO. Silibinin caused ROS generation and its effect was inhibited by calpain inhibitor, the general PKC inhibitor GF 109203X, the specific PKCδ inhibitor rottlerin, and catalase. Silibinin-induce cell death was blocked by calpain inhibitor and PKC inhibitors. Silibinin-induced PKCδ activation and disruption of △ψm were prevented by the calpain inhibitor. Silibinin induced AIF nuclear translocation and its effect was prevented by calpain inhibitor. Transfection of vector expressing microRNA of AIF prevented the silibinin-induced cell death.
  
  
  CONCLUSIONS
  Silibinin induces apoptotic cell death through a calpain-dependent mechanism involving PKC, ROS, and AIF nuclear translocation in U87MG human glioma cells.
authors.affiliation: Department of Oriental Medicine, Dongguk University, Kyung Ju, 780-714, Korea.
authors.forename: Ji C
authors.initials: JC
authors.lastname: Jeong
authors.forename: Won Y
authors.initials: WY
authors.lastname: Shin
authors.forename: Thae H
authors.initials: TH
authors.lastname: Kim
authors.forename: Chae H
authors.initials: CH
authors.lastname: Kwon
authors.forename: Jae H
authors.initials: JH
authors.lastname: Kim
authors.forename: Yong K
authors.initials: YK
authors.lastname: Kim
authors.forename: Ki H
authors.initials: KH
authors.lastname: Kim
mesh_terms: D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005910:Glioma / Q000378:metabolism; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077385:Silybin; D012838:Silymarin / Q000494:pharmacology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C494829:AIFM1 protein, human; D000975:Antioxidants; D051033:Apoptosis Inducing Factor; D017382:Reactive Oxygen Species; D012838:Silymarin; D051028:bcl-2-Associated X Protein; D000077385:Silybin; D011493:Protein Kinase C; D002154:Calpain