Contributions
Abstract: PB1895
Type: Publication Only
Background
Chronic myeloid leukemia (CML) is a clonal myeloproliferative disease. The high levels of Musashi2 (Msi2) is associated with down-regulation of Numb, a cell-fate determinant gene, in CML cells.
Aims
In current study, we knocked down Msi2 using RNA interference (RNAi) strategy and investigated the effects of this knock down on the expression of mTOR signaling.
Methods
Synthetic double stranded siRNAs designed to target human Msi2 were transfected to CD34 CML cells. Changes in the expression levels of Msi-2, Numb, TOR, and Bcl-2 genes 48h after transfection were evaluated by real-time PCR. Induction of apoptosis in transfected leukemic cells was determined using Annexin-PI staining and flowcytometry analysis.
Results
We found that upon Msi2 suppression, the expression levels of the cell-fate determinant Numb showed significantly increase in CD34+ CML cells. Msi2 down-regulation and subsequent increase in Numb expression levels caused reduction in expression of mTOR, as an oncogenic regulator. In addition, Msi2 down-regulation promoted cell apoptosis via the downregulation of Bcl-2 expression. We observed that Msi2 downregulation resulted in decreased cell proliferation and elevated rate of apoptiosis in CD34+ CML cells.
Conclusion
It seems that Msi2 could be an option for targeting CD34+ CML cells and its down-regulation through RNAi strategy may lead to induction of apoptosis in leukemic stem cells. This approach may open up new opportunities for leukemia therapy.
Session topic: 7. Chronic myeloid leukemia – Biology & Translational Research
Keyword(s): Apoptosis, Chronic myeloid leukemia, Proliferation, RNA interference (RNAi)
Abstract: PB1895
Type: Publication Only
Background
Chronic myeloid leukemia (CML) is a clonal myeloproliferative disease. The high levels of Musashi2 (Msi2) is associated with down-regulation of Numb, a cell-fate determinant gene, in CML cells.
Aims
In current study, we knocked down Msi2 using RNA interference (RNAi) strategy and investigated the effects of this knock down on the expression of mTOR signaling.
Methods
Synthetic double stranded siRNAs designed to target human Msi2 were transfected to CD34 CML cells. Changes in the expression levels of Msi-2, Numb, TOR, and Bcl-2 genes 48h after transfection were evaluated by real-time PCR. Induction of apoptosis in transfected leukemic cells was determined using Annexin-PI staining and flowcytometry analysis.
Results
We found that upon Msi2 suppression, the expression levels of the cell-fate determinant Numb showed significantly increase in CD34+ CML cells. Msi2 down-regulation and subsequent increase in Numb expression levels caused reduction in expression of mTOR, as an oncogenic regulator. In addition, Msi2 down-regulation promoted cell apoptosis via the downregulation of Bcl-2 expression. We observed that Msi2 downregulation resulted in decreased cell proliferation and elevated rate of apoptiosis in CD34+ CML cells.
Conclusion
It seems that Msi2 could be an option for targeting CD34+ CML cells and its down-regulation through RNAi strategy may lead to induction of apoptosis in leukemic stem cells. This approach may open up new opportunities for leukemia therapy.
Session topic: 7. Chronic myeloid leukemia – Biology & Translational Research
Keyword(s): Apoptosis, Chronic myeloid leukemia, Proliferation, RNA interference (RNAi)