[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.