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Published on: October 21, 2012
[The study of inhibitory and apoptosis of K562 cells by antisense oligodeoxynucleotides]
1Fujian Institute of Hematology, Union Hospital Affiliated to Fujian Medical University, Fuzhou 350001, China.
Aim:
To study the effect of bcr-abl fusion gene antisense phosphorothioate oligodeoxynucleotide (Aspo) on bcr-abl mRNA and apoptosis of K562 cells.
Methods:
Cells were exposed to Aspo. P210 was measured by Flow Cytometry. Cellular bcr-abl mRNA was detected by RT-PCR mediquantitative analysis. Cell apoptosis was measured by Flow Cytometry and observed by electron microscope (EM).
Results:
The P210 was down regulated or completely suppressed after 24h treatment with more than 5 micromol/L Aspo. The decrease of bcr-abl mRNA was about 45%. After incubation 48 h with 10 micromol/L Aspo. Also, 20% - 30% K562 cells were induced to apoptosis at 120 h when the cell number was 1 x 10(4)/ml at the beginning. While the cell number was 1 x 10(5)/ml, the apoptosis rate was 30% after 48 h culture and the typical morphology of apoptosis cell was observed under EM.
Conclusion:
bcr-abl Aspo could inhibit the expression of bcr-abl mRNA and P210. Also,it could induce apoptosis of K562 cells.
Insights
Antisense phosphorothioate oligodeoxynucleotide (Aspo) targeting the bcr-abl fusion gene effectively reduced bcr-abl mRNA and P210 protein levels. This inhibition induced significant apoptosis in K562 cancer cells.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Therapy
Background:
- The bcr-abl fusion gene is a hallmark of chronic myeloid leukemia (CML).
- Targeting oncogenic transcripts like bcr-abl is a promising therapeutic strategy.
- K562 cells, derived from CML patients, express the bcr-abl fusion gene.
Purpose of the Study:
- To investigate the efficacy of bcr-abl antisense phosphorothioate oligodeoxynucleotide (Aspo) in downregulating bcr-abl mRNA and P210 protein.
- To evaluate the impact of Aspo on inducing apoptosis in K562 cells.
Main Methods:
- K562 cells were treated with varying concentrations of Aspo.
- bcr-abl mRNA levels were quantified using RT-PCR.
- P210 protein expression was measured by Flow Cytometry.
- Apoptosis was assessed via Flow Cytometry and electron microscopy.
Main Results:
- Aspo treatment (≥5 µmol/L for 24h) significantly reduced P210 levels, with complete suppression observed at higher concentrations.
- A ~45% decrease in bcr-abl mRNA was noted after 48h of incubation with 10 µmol/L Aspo.
- Aspo induced apoptosis in 20%-30% of K562 cells after 48-120h, with characteristic morphological changes observed under electron microscopy.
Conclusions:
- bcr-abl Aspo effectively inhibits the expression of bcr-abl mRNA and its corresponding P210 protein.
- Aspo demonstrates potent apoptosis-inducing capabilities in K562 cells, suggesting therapeutic potential.
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