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[Telomerase gene antisense oligodeoxynucleotide down-regulates telomerase activity in HL-60 cells]
1Institute of Hematology, Jinan University Medical College, Guangzhou 510632, P. R. China.
Background & Objective:
Human telomerase reverse transcriptase (hTERT) is a limiting component for telomerase activity. Telomerase inhibitor targeting telomerase RNA could arrest growth of tumor cells. This study was designed to explore the effect of hTERT gene antisense phosphorothioate oligodeoxynucleotide (ASODN) on telomerase activity in HL-60 cells.
Methods:
Telomeric repeat amplification protocol was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence using fluorescence isothiocyanate label respectively.
Results:
RT-PCR and immunofluorescence assay showed that the expression levels of hTERT mRNA and protein from HL-60 cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and SODN-treated cells. Telomerase activity decreased when HL-60 cells were treated with ASODN for 48 h. Telomerase activity of HL-60 cells was significantly inhibited when treated with ASODN for 72h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells.
Conclusion:
Telomerase activity could be down-regulated by hTERT ASODN in HL-60 cells.
Insights
Human telomerase reverse transcriptase (hTERT) antisense oligodeoxynucleotides effectively reduced telomerase activity in HL-60 cancer cells. This targeted approach shows potential for controlling tumor cell growth by inhibiting telomerase.
Area of Science:
- Molecular Biology
- Cancer Research
- Oligonucleotide Therapeutics
Context:
- Telomerase activity is crucial for cancer cell proliferation and survival.
- Human telomerase reverse transcriptase (hTERT) is a key component of telomerase.
- Targeting telomerase offers a potential strategy for cancer treatment.
Purpose:
- To investigate the effect of hTERT gene antisense phosphorothioate oligodeoxynucleotide (ASODN) on telomerase activity in HL-60 cells.
- To evaluate the impact of hTERT ASODN on hTERT mRNA and protein expression levels.
- To assess the inhibition of telomerase activity by hTERT ASODN.
Summary:
- hTERT ASODN treatment led to decreased hTERT mRNA and protein expression in HL-60 cells over time.
- Telomerase activity was reduced after 48 hours and significantly inhibited after 72 hours of hTERT ASODN treatment.
- No significant changes in hTERT expression or telomerase activity were observed in control or SODN-treated cells.
Impact:
- hTERT ASODN demonstrates efficacy in down-regulating telomerase activity in HL-60 cells.
- This study supports the potential of hTERT ASODN as a therapeutic agent for cancers characterized by high telomerase activity.
- The findings contribute to the development of novel anti-cancer strategies targeting telomerase.