Objective: To investigate the effect of targeted inhibition of CD47 gene expression on stem cell clearance in acute myeloid leukemia.
Methods: After the lentiviral CD47-siRNA was transfected into acute myelogenous leukemia stem cells (LSCs), the proliferative status of acute myelogenous LSCs was detected by cell counting kit-8, and the apoptosis of stem cells of acute myeloid leukemia was detected by annexin/propidium iodide flow assays. The expression of Bcl-2, Bcl-xl, MCL-1, PIK3p110β, and interleukin (IL)-3 in acute myeloid LSCs was detected by Western blot analysis and the activity of protein phosphatase 2A (PP2A) and the protein content of CD96 and CD90 were measured by enzyme-linked immunosorbent assay kits.
Results: After transfection of the lentivirus CD47-siRNA into acute myeloid LSCs, compared with the empty vector transfection group (control group), the cell viability of the CD47-siRNA transfection group was decreased, and the apoptosis rate was increased. Furthermore, the antiapoptotic protein Bcl-2, Bcl-xl, and MCL-1 and the content of IL-3 protein, CD96, and CD90 was decreased, whereas the activity of PIK3p110β and PP2A protein was increased.
Conclusion: Targeted inhibition of CD47 could inhibit the proliferation of myeloid LSCs, promote apoptosis, mobilize the cells into the cell cycle, and reduce the high expression of immune proteins on the cell surface, therefore providing a theoretical basis for the elimination and eradication of LSCs.
Keywords: CD47; apoptosis; cell cycle; gene silence; leukemia stem cells; proliferation.
© 2018 Wiley Periodicals, Inc.