Nicotine induces apoptosis in TM3 mouse Leydig cells.
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.
Kim
Khae-Hawn
KH
Department of Urology, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.
Joo
Kwan-Joong
KJ
Park
Heung-Jae
HJ
Kwon
Chil-Hun
CH
Jang
Mi-Hyeon
MH
Kim
Chang-Ju
CJ
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
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