title: Specific features of apoptosis in human lens epithelial cells induced by mitomycin C in vitro.
abstract: BACKGROUND
  Posterior capsule opacification (PCO) is still one of the major complications following modern cataract surgery. Several attempts have been made to find an appropriate therapeutic concept to significantly lower the rate of PCO. Here, we wanted to focus on the antimetabolic strategy, reducing PCO by using mitomycin C, further characterizing the pathway of apoptosis in human lens epithelial cells (hLECs).
  
  
  METHODS
  Human lens epithelial cells were obtained from anterior lens capsules during cataract surgery. The expression of Fas, TRAMP, TRAIL-R1-R4, Apo-3L and TRAIL mRNA was investigated by means of RT-PCR using specific primers. For investigations on bcl-2, bax, p53 and the active form of caspase 3, cell cultures of hLECs were pretreated with mitomycin C and processed for immunocytochemistry thereafter.
  
  
  RESULTS
  We detected the expression of the receptors Fas, TRAMP, TRAIL-R2 and TRAIL-R3 in hLECs. We further obtained evidence of the upregulation of the intracellular apoptotic signalling cascade, represented by bcl-2 and bax, the transcription factor p53 and the active form of caspase 3, after pretreatment with mitomycin C.
  
  
  CONCLUSIONS
  We demonstrated the presence of the apoptosis-receptor system in hLECs. Furthermore, we demonstrated the possibility of the induction of key proteins of the apoptotic signalling cascade in these cells by the antimetabolic drug mitomycin C. This could have important implications on the strategies regarding both the prevention and the treatment of PCO after cataract surgery.
authors.affiliation: University Eye Clinic, University of Cologne, Germany. j.jordan@uni-koeln.de
authors.forename: J F
authors.initials: JF
authors.lastname: Jordan
authors.forename: N
authors.initials: N
authors.lastname: Kociok
authors.forename: S
authors.initials: S
authors.lastname: Grisanti
authors.forename: P C
authors.initials: PC
authors.lastname: Jacobi
authors.forename: J M
authors.initials: JM
authors.lastname: Esser
authors.forename: T T
authors.initials: TT
authors.lastname: Luther
authors.forename: G K
authors.initials: GK
authors.lastname: Krieglstein
authors.forename: P
authors.initials: P
authors.lastname: Esser
mesh_terms: D000963:Antimetabolites / Q000494:pharmacology*; D017209:Apoptosis / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007908:Lens, Crystalline / Q000187:drug effects* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D016685:Mitomycin / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics; D053219:Receptors, Tumor Necrosis Factor, Member 25; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein; D019014:fas Receptor / Q000235:genetics
chemical_list: D000963:Antimetabolites; D051017:Apoptosis Regulatory Proteins; C494826:BAX protein, human; D011508:Chondroitin Sulfate Proteoglycans; D017931:DNA Primers; C082019:DPT protein, human; D016326:Extracellular Matrix Proteins; D008562:Membrane Glycoproteins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D018124:Receptors, Tumor Necrosis Factor; D053219:Receptors, Tumor Necrosis Factor, Member 25; D053221:TNF-Related Apoptosis-Inducing Ligand; C506033:TNFRSF25 protein, human; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D019014:fas Receptor; D016685:Mitomycin; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases