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Human hepatoma cell telomerase activity inhibition and cell cycle modulation by its RNA component antisense
R G Zhang1, X W Wang, J H Yuan
1Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
Aim:
To investigate the effects of human telomerase RNA component antisense oligodeoxyribonucleotides on telomerase activity of human hepatoma cells and their effects on cell cycle distribution.
Methods:
Modified telomeric repeat amplification protocol was used to detect telomerase activity. Cell cycle was analyzed by flow cytometer.
Results:
Telomerase activity was detected in all of four human hepatoma cell lines but absent in normal liver cells. Antisense oligomers to human telomerase RNA component (hTR) inhibited telomerase activity of BEL-7404 human hepatoma cells markedly in vitro. After in vitro treatment with antisense oligomers for 96 h, cell cycle of BEL-7404 human hepatoma cells was mainly arrested at G2/M phase.
Conclusion:
Antisense oligomers to hTR inhibited telomerase activity of BEL-7404 human hepatoma cells in vitro and resulted in cell cycle arrest at G2/M phase.
Insights
Antisense oligodeoxyribonucleotides targeting the human telomerase RNA component (hTR) effectively inhibited telomerase activity in human hepatoma cells. This inhibition led to cell cycle arrest at the G2/M phase, offering a potential therapeutic strategy.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Telomerase is crucial for maintaining telomere length and is often reactivated in cancer cells, including hepatoma.
- Human telomerase RNA component (hTR) is essential for telomerase function.
- Targeting telomerase presents a potential strategy for cancer therapy.
Purpose of the Study:
- To evaluate the impact of antisense oligodeoxyribonucleotides against hTR on telomerase activity in human hepatoma cells.
- To determine the effects of these antisense oligodeoxyribonucleotides on hepatoma cell cycle progression.
Main Methods:
- Telomerase activity was assessed using a modified telomeric repeat amplification protocol.
- Cell cycle distribution was analyzed via flow cytometry.
- Human hepatoma cell lines (BEL-7404) were treated with antisense oligodeoxyribonucleotides targeting hTR in vitro.
Main Results:
- Telomerase activity was present in all tested human hepatoma cell lines but not in normal liver cells.
- Antisense oligodeoxyribonucleotides against hTR significantly inhibited telomerase activity in BEL-7404 cells.
- Treatment with antisense oligodeoxyribonucleotides for 96 hours resulted in cell cycle arrest predominantly at the G2/M phase in BEL-7404 cells.
Conclusions:
- Antisense oligodeoxyribonucleotides targeting the human telomerase RNA component (hTR) demonstrate potent inhibition of telomerase activity in human hepatoma cells.
- Inhibition of telomerase by these antisense oligodeoxyribonucleotides leads to G2/M phase cell cycle arrest, suggesting a role in controlling hepatoma cell proliferation.