hTERT antisense oligonucleotides enhance apoptosis induced by cisplatin in HeLa cell lines / 中国药理学通报

ABSTRACT

Aim To investigate the effects of human telomerase reverse transcriptase(hTERT)antisense phosphorothioate oligodeoxynucleotide(ASODN)on telomerase activity and apoptosis induced by crisplatin(DDP)in HeLa cell lines.Methods HeLa cells were collected after transfection of 0.05,0.1,0.2 ?mol?L-1of ASODN and the total DNA was extracted.Telomeric repeat amplification protocol(TRAP)-AgNO3 stain was used to detect the activity of telomerase.Apoptosis of HeLa cells was detected by AO staining morphological observation and flow cytometry(FCM)analysis technology.Results The telomerase activity of HeLa cells was expressed in high level,which was significantly reduced dose-dependently and time-dependently after treatment with ASODN of hTERT.Two days after transfection of ASODN of 0.05 ?mol?L-1,0.1 ?mol?L-1,0.2 ?mol?L-1,the telomerase activity decreased by 21.77%,52.42% and 71.10% respectively.After treatment with ASODN for 24,48 h and 72 h,the telomerase activity decreased by 12.48%,38.27% and 71.10% respectively.In morphological observation using acridine orange(AO)staining method,HeLa cells displayed typical apoptotic figure after treated with DDP plus ASODN of hTERT.The percentage of apoptosis of HeLa cells treated with DDP of 1.5 mg?L-1,3.0 mg?L-1,for 48 hours after 24 hours exposure to ASODN of 0.2 ?mol?L-1(50.35%,29.67%)was significantly higher than that treated with DDP alone(19.67%,11.38%,P