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Published on: September 25, 2011
Effects of phosphorothioate anti-sense oligodeoxynucleotides on colorectal cancer cell growth and telomerase activity
Xi-Shan Wang1, Kuan Wang, Xue Li
1Department of Abdominal Surgery, Tumour Hospital of Harbin Medical University, Harbin 150040, Heilongjiang Province, China. 1966120808@sina.com
Aim:
To investigate the inhibitory effect of phosphorothioate anti-sense oligodeoxynucleotides (PASODN) on colorectal cancer LS-174T cells in vitro and the mechanism of inhibition of telomerase activity in these cells.
Methods:
PASODN were used to infect LS-174T cells and block human telomerase RNA (hTR) through anti-sense technology. The inhibitory effect of PASODN was evaluated by colony-forming inhibition assay and growth curve. Changes of telomerase activity in LS-174T cells were detected by polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA), and the level of apoptosis was analyzed by flow cytometry (FCM) assay.
Results:
PASODN showed a dose and time-dependent inhibition of cell proliferation. The optimal dosage of PASODN was 10 mumol/L. The colony-forming efficiency was 10.3% in PASODN group after 10 d, whereas that in phosphorothioate mis-sense oligodeoxynucleotides (PMSODN) group with the same concentration and in PBS group (blank control) was 49.1% and 50.7%, respectively. PCR-ELISA results indicated that telomerase activity in the PASODN group was obviously inhibited in comparison with in the control groups (P<0.01, t = 3.317 and 3.241, t0.01(20) = 2.845). Meanwhile, before the number of cells was decreased, the morphological changes were observed in the cells of PASODN group. The cells in PASODN group showed the apoptotic peak at 72 h after infection, whereas the control group did not show.
Conclusion:
Specific sequence oligonucleotides can inhibit telomerase activity and lead to cell apoptosis, suggesting a novel treatment strategy for malignant tumors induced by telomerase.
Insights
Phosphorothioate anti-sense oligodeoxynucleotides (PASODN) effectively inhibit colorectal cancer cell proliferation by blocking human telomerase RNA. This targeted approach induces cancer cell apoptosis, offering a promising new therapeutic strategy.
Area of Science:
- Molecular Biology
- Cancer Research
- Oligonucleotide Therapeutics
Background:
- Telomerase is a key enzyme in cancer cell immortalization and proliferation.
- Targeting telomerase offers a potential strategy for cancer treatment.
- Phosphorothioate anti-sense oligodeoxynucleotides (PASODN) can be designed to inhibit specific RNA targets.
Purpose of the Study:
- To evaluate the in vitro inhibitory effect of PASODN on colorectal cancer LS-174T cells.
- To investigate the mechanism by which PASODN inhibits telomerase activity.
- To assess the potential of PASODN as a novel anti-cancer therapeutic strategy.
Main Methods:
- LS-174T cells were treated with PASODN designed to block human telomerase RNA (hTR).
- Cell proliferation was assessed using colony-forming inhibition assays and growth curves.
- Telomerase activity was measured by PCR-ELISA, and apoptosis was analyzed by flow cytometry.
Main Results:
- PASODN demonstrated a dose- and time-dependent inhibition of LS-174T cell proliferation.
- Optimal PASODN dosage (10 μmol/L) significantly reduced colony formation compared to controls.
- PASODN treatment markedly inhibited telomerase activity and induced significant apoptosis in cancer cells.
Conclusions:
- Specific sequence PASODN effectively inhibits telomerase activity in colorectal cancer cells.
- Inhibition of telomerase leads to cancer cell apoptosis, validating this as a therapeutic approach.
- Oligonucleotide-based telomerase inhibition represents a novel strategy for treating telomerase-dependent malignancies.
