[Effect of antisense oligonucleotide against telomerase in rat spermatogonia and its reactivity to cytokine].
OBJECTIVE
To study the change of telomerase activity in rat spermatogonia when the telomerase RNA was enclosed, and reactivity of the change to cytokine(SCF, TGF-beta 1).


METHODS
The antisense oligonucleotides(PS-ASON) of telomerase was transfected into proliferating spermatogonia in vitro with the liposomes as the vector. Then the cytokine, stem cell factor (SCF) or transforming growth factor-beta 1(TGF-beta 1), was added. The proliferative activity of the spermatogonia was determined before and after the inhibition by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]assay. The change of telomerase activity was detected by telomeric repeat amplification protocol (TRAP).


RESULTS
1 mumol/L PS-ASON obviously downregulated the telomerase activity and inhibited spermatogonia proliferation. When the inhibition was over, the activity recovered to some extent(P < 0.01). Growth factors can regulate the spermatogonia after inhibition, SCF may improve the activity of telomerase and the proliferation of spermatogonia. Adversely, TGF-beta 1 may inhibit the recovery of telomerase activity.


CONCLUSIONS
To inhibit spermatogonia telomerase activity antisensely can limit the proliferation of spermatogonia efficiently, which was regulated by cytokine. This method might be a new and efficient way in male birth control.
Ping
Hao
H
Department of Urology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, China.
Chen
Xiao-Chun
XC
Ye
Zhe-Wei
ZW
Zhang
Yan
Y
Zhang
Run-Qing
RQ
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D003272:Contraceptive Agents, Male / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D008297:Male; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D016601:RNA-Binding Proteins / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013093:Spermatogonia / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D019089:Stem Cell Factor / Q000378:metabolism; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis; D014162:Transfection
D001426:Bacterial Proteins; D003272:Contraceptive Agents, Male; D016207:Cytokines; C079092:MtrB protein, Bacteria; D016376:Oligonucleotides, Antisense; D016601:RNA-Binding Proteins; D019089:Stem Cell Factor; D014157:Transcription Factors; D019098:Telomerase