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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。
Objective To study the effects of a novel small molecule tyrosine kinase inhibitor Apatinib on the proliferation and apoptosis of stem and progenitor cells of acute myeloid leukemia (AML) and its related molecular mechanisms. Methods CCK8 was used to detect the inhibitory effect of different concentrations of Apatinib on the proliferation of kg1α cells. The apoptosis of kg1α cells and primary CD34 + AML stem cells induced by different concentrations of Apatinib was detected by Annexin V / PI method. The expression of PI3K / AKT pathway related proteins (AKT, Raf and PTEN) expression changes. Results Apatinib (2.5, 5, 10, 20, 40 μmol·L -1) had significant inhibitory effects on the proliferation of kg 1α cells at 48 h and 72 h in a dose- and time-dependent manner. Compared with the control The differences between the two groups were statistically significant (P <0.05 or P <0.01). The results of Annexin V / PI assay showed that apatinib at different concentrations significantly induced apoptosis of kg1α, and the apoptotic rates at 48 h and 72 h were significantly different from those of the control group (P < 0.01). The effect of different concentrations of Apatinib on the proliferation of 7 primary AML stem cells after 48 h was significant (P <0.01). The results of Western blot showed that Apatinib treatment of kg 1α cells for 48 h After AKT / p-AKT, p-Raf expression decreased, while p-PTEN expression increased. Conclusion Apatinib can inhibit the proliferation of AML-like progenitor cells and induce the apoptosis of both K1 and Aβ stem cells in a concentration-and time-dependent manner. The mechanism may be through interfering with PI3K / AKT pathway to achieve.