title: Nicotine induces apoptosis in TM3 mouse Leydig cells.
abstract: OBJECTIVE
  To investigate whether nicotine-induced testicular toxicity involves the induction of apoptosis in Leydig cells.
  
  
  METHODS
  Cell study using cells of the TM3 cell line derived from mouse Leydig cells.
  
  
  METHODS
  Academic research laboratory.
  
  
  METHODS
  Morphological and biochemical analyses for the detection of apoptosis.
  
  
  METHODS
  The effect of nicotine on the occurrence of apoptosis was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6-diamidino-2-phenylindole staining, terminal deoxynuclotidyl transferase-mediated dUTP nick end labeling assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction, caspase-3 enzyme assay, and Western blot analysis. Nicotine treatment exhibits several features of apoptosis in mouse Leydig cells.
  
  
  RESULTS
  TM3 cells treated with nicotine exhibit several features of apoptosis. It was also shown that nicotine increases the mRNA level of bax and decreases that of bcl-2. In addition, nicotine enhanced the expression of the activated form of caspase-3 and caspase-3 enzyme activity.
  
  
  CONCLUSIONS
  Nicotine appears to activate specific intracellular death-related pathways, probably by bax-dependent activation of caspase-3, inducing apoptosis in Leydig cells. Thus, nicotine-induced apoptosis of Leydig cells might be one of the important mechanisms behind nicotine-related urogenital disorders in men.
authors.affiliation: Department of Urology, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.
authors.forename: Khae-Hawn
authors.initials: KH
authors.lastname: Kim
authors.forename: Kwan-Joong
authors.initials: KJ
authors.lastname: Joo
authors.forename: Heung-Jae
authors.initials: HJ
authors.lastname: Park
authors.forename: Chil-Hun
authors.initials: CH
authors.lastname: Kwon
authors.forename: Mi-Hyeon
authors.initials: MH
authors.lastname: Jang
authors.forename: Chang-Ju
authors.initials: CJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects*; D007985:Leydig Cells / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D008297:Male; D051379:Mice; D009538:Nicotine / Q000633:toxicity*; D018722:Nicotinic Agonists / Q000633:toxicity*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D051028:bcl-2-Associated X Protein
chemical_list: C494827:Bax protein, mouse; D018722:Nicotinic Agonists; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; D009538:Nicotine; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases