[Combination of antisense human telomerase RNA and antisense human telomerase catalytic subunit inhibits cervical cancer Hela cells growth].
OBJECTIVE
To investigate the inhibitory effect of combination of antisense human telomerase RNA (anti-hTR) and antisense human telomerase catalytic subunit (anti-hTERT) on proliferation of cervical cancer Hela cell line cultured in vitro.


METHODS
Cervical cancer Hela cells were transfected by anti-hTR, anti-hTERT, anti-hTR + anti-hTERT, sense-hTR, sense-hTERT with Oligofectamin reagent. The proliferation activity of cervical cancer Hela cells was determined using methyl thiazolyl tetrazolium (MTT). The activity of telomerase was tested by polymerase chain reaction-telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (PCR-TRAP-ELISA). Cell morphologies were observed under fluorescence microscope with acridine orange staining. Apoptosis and cell cycle were examined by flow cytometer method (FCM).


RESULTS
The inhibitory rates on proliferation and apoptotic rates of Hela cells transfected by antisense oligonucleotides were significantly higher compared with those of control, sense oligonucleotides and Oligofectamin (all P < 0.01). Meanwhile, the inhibitory rates on proliferation and apoptotic rates of Hela cells transfected by anti-hTR + anti-hTERT were significantly higher compared with those transfected by anti-hTR or anti-hTERT alone (P < 0.01); telomerase activity was significantly decreased (P < 0.01). Transfected by antisense oligonucleotides at 0.2 micromol/L for 3 d, inhibitory rates on proliferation, telomerase activity (relative to control), apoptotic rate of Hela cells was 69.3%, 19.6%, 28.6% respectively (Q = 0.867, 0.919, 1.075). Percentages of G(0)/G(1) phase cells were increased.


CONCLUSIONS
Telomerase anti-hTR and anti-hTERT inhibit Hela cell growth probably through suppressing telomerase activity, inducing apoptosis and blocking cell cycle. Combination of anti-hTR and anti-hTERT has synergistic action on cervical cancer Hela cells.
Lei
Li-xiu
LX
Department of Obstetrics and Gynecology, Nanhua University First Hospital, Hengyang 421001, China.
Chen
Zhong-dong
ZD
D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D004268:DNA-Binding Proteins; D004357:Drug Synergism; D005260:Female; D006367:HeLa Cells; D006801:Humans; D016372:RNA, Antisense / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000201:enzymology / Q000235:genetics / Q000473:pathology*
D004268:DNA-Binding Proteins; D016372:RNA, Antisense; D019098:Telomerase