Morphological Laboratory, Taishan Medical College, Taian, Shandong, China.
Department of Pathology, Taishan Medical College, Taian, Shandong, China.
J Cell Biochem. 2019 Jun;120(6):10303-10309. doi: 10.1002/jcb.28314. Epub 2018 Dec 19.
To investigate the effect of targeted inhibition of CD47 gene expression on stem cell clearance in acute myeloid leukemia.
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.
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.
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.
研究靶向抑制 CD47 基因表达对急性髓系白血病干细胞清除的影响。
将慢病毒 CD47-siRNA 转染入急性髓系白血病干细胞(LSCs)后,用细胞计数试剂盒-8 检测急性髓系 LSCs 的增殖状态,用 Annexin/碘化丙啶流式检测急性髓性白血病干细胞的凋亡。Western blot 分析检测急性髓系 LSCs 中 Bcl-2、Bcl-xl、MCL-1、PIK3p110β 和白细胞介素(IL)-3 的表达,酶联免疫吸附试验试剂盒检测蛋白磷酸酶 2A(PP2A)的活性和 CD96 和 CD90 的蛋白含量。
慢病毒 CD47-siRNA 转染急性髓系 LSCs 后,与空载体转染组(对照组)相比,CD47-siRNA 转染组细胞活力降低,凋亡率增加。此外,抗凋亡蛋白 Bcl-2、Bcl-xl 和 MCL-1 以及 IL-3 蛋白、CD96 和 CD90 的含量降低,而 PIK3p110β 和 PP2A 蛋白的活性增加。
靶向抑制 CD47 可抑制髓系 LSCs 的增殖,促进凋亡,使细胞进入细胞周期,并降低细胞表面免疫蛋白的高表达,为清除和根除 LSCs 提供了理论依据。