白杨素对人白血病细胞柔红霉素化疗敏感性及PI3K/Akt通路的影响  

Effects of Chrysin on the Chemosensitivity of Daunorubicin in Human Leukemia Cells by Inhabiting PI3K/Akt Pathway

作　　者：曹福英 张春霞 孙孟炎[2] 桑玉旗[1] 夏建胜[1] Cao Fuying;Zhang Chunxia;Sun Mengyan;Sang Yuqi;Xia Jiansheng(Department of Hematology,Heze Municipal Hospital,Shandong Heze 274000,China;Department of Clinical Laboratory,Heze Municipal Hospital,Shandong Heze 274000,China)

机构地区：[1]菏泽市立医院血液科,山东菏泽274000 [2]菏泽市立医院检验科,山东菏泽274000

出　　处：《中国药师》2020年第6期1037-1041,1047,共6页China Pharmacist

摘　　要：目的:研究白杨素联合柔红霉素诱导体外培养的急性粒细胞性白血病细胞株(HL-60)的细胞凋亡作用,通过其对磷酸肌醇-3激酶/蛋白激酶B(PI3K/Akt)信号通路的活化,增加HL-60对柔红霉素的敏感性。方法:用MTT法测定对照组、白杨素组(20μmol·L^-1)、柔红霉素组(0.5μg·ml^-1)和白杨素(20μmol·L^-1)+柔红霉素(0.5μg·ml^-1)组对HL-60细胞的抑制率,观察各组HL-60细胞形态变化,并用吖啶橙和溴化乙锭(AO/EB)双荧光法观察细胞凋亡情况;用逆转录-聚合酶链反应(RT-PCR)方法检测白杨素与柔红霉素联用对HL-60细胞PI3K相关基因S期激酶相关蛋白2(Skp-2)、抑癌基因P27、B淋巴细胞瘤-2基因(Bcl-2)和Bcl-2相关X蛋白(Bax)表达的影响;用蛋白质免疫印迹法(Western Blotting)方法检测白杨素与柔红霉素联用对HL-60细胞PI3K相关蛋白Skp-2、P27、Bcl-2和Bax表达的影响。结果:白杨素与柔红霉素联用能显著增强HL60对柔红霉素的化疗名感性,降低HL60细胞存活率,增加细胞凋亡,下调Skp-2、Bcl-2 mRNA及蛋白表达,上调P27 mRNA及蛋白表达,与单药组比较差异有统计学意义(P<0.05);而Bax mRNA及蛋白表达变化不明显(P>0.05)。结论:白杨素可以有效靶向诱导HL-60肿瘤细胞凋亡,并通过PI3K/Akt信号通路增加HL-60对柔红霉素的化疗敏感性。Objective:To study the apoptosis of acute myeloid leukemia cell line(HL-60)induced by chrysin and daunorubicin in vitro,and increase the sensitivity of HL-60 to daunorubicin through its activation of phosphoinositide-3 kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods:MTT method was used to determine the inhibition rate of HL-60 cells in the control group,chrysin group(20μmol·L^-1),daunorubicin group(0.5μg·ml^-1)and chrysin(20μmol·L^-1)+daunorubicin(0.5μg·ml^-1)group;the morphological changes of HL-60 cells in each group were observed;the cell apoptosis was observed with acridine orange and ethidium bromide(AO/EB)double fluorescence method;the reverse transcription-polymerase chain reaction(RT-PCR)method was used to detect the combined use of chrysin and daunorubicin on the expressions of PI3K-related gene S-phase kinase-related protein 2(Skp-2),tumor suppressor gene P27,B-lymphoma-2 gene(Bcl-2)and Bcl-2 related X protein(Bax);Western blotting was used to detect the effects of chrysin and daunorubicin on the expressions of PI3K-related proteins Skp-2,P27,Bcl-2 and Bax in HL-60 cells.Results:The combination of chrysin and daunorubicin could significantly enhance the sensitivity of HL60 to daunorubicin,reduce the survival rate of HL60 cells,increase apoptosis,down-regulate the expressions of Skp-2,Bcl-2 mRNA and protein,up-regulate the expressions of P27 mRNA and protein,and there was a statistically significant difference compared with the single drug group(P<0.05).But the expressions of Bax mRNA and protein did not change significantly(P>0.05).Conclusion:Chrysin can effectively targeting induce the apoptosis of HL-60 tumor cells and increase the chemosensitivity of HL-60 to daunorubicin through PI3K/Akt signaling pathway.

关 键 词：白杨素 白血病 柔红霉素 HL-60细胞株 磷酸肌醇-3激酶/蛋白激酶B 

分 类 号：R965.1[医药卫生—药理学]