Antisense Sp1 oligodeoxynucleotide decreases telomerase activity by inhibiting hTERT mRNA expression in Jurkat T cells.
OBJECTIVE
To study the effects of transcriptional factor Sp1 antisense oligodeoxynucleotide (ODN) on telomerase activity and human telomerase reverse transcriptase (hTERT) expression.


METHODS
Antisense oligodeoxynucleotide (ODN) was designed to inhibit Sp1 expression and transferred to Jurkat T cells by lipofectamin. Telomerase PCR-ELISA was used to detect telomerase activity. RT-PCR analysis was used to assess the mRNA expression of Sp1 and hTERT, and Western blot was used to analyze the levels of Sp1 protein.


RESULTS
Treatment of Jurkat T cells with Sp1 antisense ODN (1 micromol/L) dramatically reduced Sp1 mRNA and protein levels. The inhibition rate was 44.8 % (P <0.05) and 57 % (P <0.01), respectively. Following the transcriptional factor Sp1 functionally altering, hTERT mRNA expression were suppressed with a 43.7 % inhibition rate (P <0.01). A dose-dependent inhibition of telomerase activity by antisense Sp1 ODN was also discovered. From 0.25 to 2.0 micromol/L, telomerase activity was reduced from 27.1 % to 64.6 %.


CONCLUSIONS
Antisense Sp1 ODN decreases telomerase activity by inhibiting hTERT mRNA expression in Jurkat T cells.
Pang
Jian-Xin
JX
Institute of Pharmaceutic Sciences, First Military Medical University, Guangzhou 510515, China. pjx@fimmu.com
Cheng
Xi-Yuan
XY
Xu
Wei
W
Wu
Shu-Guang
SG
D016373:DNA, Antisense / Q000494:pharmacology*; D004268:DNA-Binding Proteins; D006801:Humans; D019169:Jurkat Cells / Q000201:enzymology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000187:drug effects; D016329:Sp1 Transcription Factor / Q000037:antagonists & inhibitors* / Q000235:genetics; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*
D016373:DNA, Antisense; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; D019098:Telomerase