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目的观察齐墩果酸(OA)诱导人急性髓系白血病M2型细胞株Kasumi-1凋亡及对AML1-ETO表达的影响。方法用不同浓度OA作用于人急性髓系白血病M2型Kasumi-1细胞株,四氮唑蓝(MTT)法检测细胞增殖抑制率;Annexin V/PI双染色后,流式细胞仪检测细胞凋亡;Western blot法检测AML1-ETO蛋白表达水平;荧光原位杂交(Fish)检测融合基因AML1-ETO的变化。结果 OA以时间和剂量依赖方式抑制Kasumi-1细胞的增殖,24,48,72 h的IC50分别约为0.58,0.41,0.35μmol.L-1。不同浓度(0.4,0.6,0.9μmol.L-1)OA作用24 h后,细胞凋亡率明显增加,AML1-ETO蛋白表达下调。结论 OA对Kasumi-1细胞的抑制增殖和诱导凋亡作用可能与AML1-ETO的抑制有关。
Objective To observe the effect of oleanolic acid (OA) on the apoptosis and the expression of AML1-ETO in human M2 cell line Kasumi-1 induced by acute myeloid leukemia. Methods Kasumi-1 cell line of human acute myeloid leukemia type M2 was treated with different concentrations of OA, and the cell proliferation inhibition rate was detected by MTT method. After the cells were stained with Annexin V / PI double staining, apoptosis was detected by flow cytometry ; Western blot was used to detect the expression of AML1-ETO protein; Fluorescence in situ hybridization (Fish) was used to detect the change of fusion gene AML1-ETO. Results OA inhibited the proliferation of Kasumi-1 cells in a time-and dose-dependent manner. The IC50 values at 24, 48 and 72 h were about 0.58, 0.41 and 0.35 μmol·L-1, respectively. After treated with different concentrations of 0.4, 0.6, 0.9 μmol·L-1 for 24 h, the apoptosis rate was significantly increased and the expression of AML1-ETO protein was down-regulated. Conclusion The inhibitory effect of OA on the proliferation and apoptosis of Kasumi-1 cells may be related to the inhibition of AML1-ETO.