Apatinib对急性髓系白血病干祖细胞样细胞株kg1α的杀伤作用及其分子机制

    Apatinib Inhibits Proliferation and Induces Apoptosis of Acute Myeloid Leukemia Stem/progenitor Like Cell Line (kg1α cells) and Its Mechanism

    • 摘要: 目的 研究新型小分子酪氨酸激酶抑制剂Apatinib对急性髓系白血病(acute myeloid leukemia,AML)干祖细胞增殖和凋亡的影响及其相关分子机制。方法 CCK8法检测不同浓度Apatinib对kg1α细胞的增殖抑制作用,Annexin V/PI法检测不同浓度Apatinib诱导kg1α细胞和原代CD34+AML干细胞的凋亡情况,Western blot法检测Apatinib处理kg1α细胞后PI3K/AKT通路相关蛋白(AKT、Raf和PTEN)的表达变化。结果 不同浓度的Apatinib (2.5,5,10,20,40µmol·L-1)作用48 h和72 h后对kg1α细胞均具有显著的增殖抑制作用,呈浓度和时间依赖性,与对照组相比差异均具有统计学意义(P<0.05或P<0.01)。Annexin V/PI检测细胞凋亡的结果显示,不同浓度apatinib对kg1α具有显著的诱导凋亡作用,作用48 h和72 h后的凋亡率和对照组相比差异均有统计学意义(P<0.01)。不同浓度Apatinib作用48 h后对7例原代AML干细胞均具有显著的杀伤作用,与对照组相比差异具有显著的统计学意义(P<0.01);Western blot结果显示,Apatinib处理kg1α细胞48 h后AKT/p-AKT、p-Raf表达降低,而p-PTEN表达增加。结论 Apatinib可抑制AML干祖细胞样细胞kg1α细胞的增殖,且可诱导kg1α细胞和原代CD34+AML干祖细胞的凋亡,均呈浓度和时间依赖性,其作用机制可能是通过干扰PI3K/AKT通路实现的。

       

      Abstract: OBJECTIVE To investigate the effect of Apatinib, a small-molecule vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor, on the proliferation and apoptosis of acute myeloid leukemia(AML) stem/progenitor cells and its molecule mechanism.METHODS The kg1α cells and primary CD34+ AML stem cells were treated with a serial of concentrations of Apatinib for 48 h and 72 h, the inhibitory ratio was measured by CCK8 assay, the apoptosis percent was measured by flow cytometry. Western bolt was used to analyzed AKT/p-AKT, p-Raf and p-PTEN expression after treatment with 0, 10, 20 μmol·L-1 Apatinib in kg1α cells.RESULTS After treatment with a serial of Apatinib (2.5, 5, 10, 20, 40 μmol·L-1) on AML stem-like cell line(kg1α) for 48 h and 72 h, the cell proliferation were significantly inhibited in a dose-and time-dependent mode. All the differences had statistical significance compared with control group. The results of Annexin V/PI showed that various concentration of Apatinib induced significantly apoptosis on kg1α cells. After 48 h and 72 h, all the apoptosis percentage were significantly higher than control group(P<0.01); Apatinib significantly induced apoptosis of 7 cases primary AML stem cell, the difference has statistical significance; Western blot results indicated decrease the expression of AKT/p-AKT and p-Raf, however, upregulated the level of p-PTEN.CONCLUSION Apatinib can inhibit the proliferation of AML stem/progenitor like cell line(kg1α cells) and also induce the apoptosis of kg1α and primary CD34+ AML stem cells. Its mechanism may be related with the PI3K/AKT signal pathway.

       

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