白皮杉醇扼制急性髓系白血病细胞恶性生物学特性的机制研究

The Mechanisms of Piceatannol in Inhibiting the Malignant Biological Characteristics of Acute Myeloid Leukemia Cells

目的:探讨白皮杉醇对急性髓系白血病(AML)细胞恶性生物学特性的影响及其分子机制。方法:使用不同浓度白皮杉醇干预HL60、U937及HL60/ADR、U937/ADR细胞,CCK-8法检测细胞增殖;Annexin V/PI双染流式细胞术检测细胞凋亡;Western blot检测凋亡、自噬及相关信号通路蛋白的表达;实时荧光定量聚合酶链式反应(qRT-PCR)检测耐药株中各耐药基因的表达变化。结果:白皮杉醇可抑制HL60、U937细胞活性,并诱导其凋亡。当白皮杉醇干预AML细胞24 h后,自噬标记物LC3-II/LC3-I比值随药物浓度增加而升高(r=0.672,r=0.549),当白皮杉醇干预48 h后,可下调AML细胞中Bcl-2蛋白的表达并水解活化caspase-3,同时抑制Akt/NF-κB信号通路活化水平以诱发AML细胞程序性死亡。白皮杉醇干预AML耐药株后亦可下调MRP1的表达,能逐渐削弱白血病耐药株的化疗抗性,但对BCRP表达的影响微弱,对MDR1基本无影响。结论:白皮杉醇能抑制AML细胞的增殖活性,并诱导其发生程序性死亡,其机制可能与抑制Akt/NF-κB信号通路激活、水解活化caspase-3并下调Bcl-2蛋白表达以及使部分耐药基因表达受抑等有关。

Objective:To explore the effect and molecular mechanism of Piceatannol on malignant biological characteristics of acute myeloid leukemia(AML)cells.Methods:HL60,U937,HL60/ADR and U937/ADR cells were treated with different concentrations of Piceatannol.CCK-8 assay was used to detect cell proliferation.Cell apoptosis was detected by flow cytometry with Annexin V/PI double staining.The protein expressions of apoptosis,autophagy and related sig naling pathways were detected by Western blot.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression changes of drug resistance genes in drug-resistant AML cell lines.Results:The activity of HL60 and U937 cells could be inhibited by Piceatannol and induced apoptosis.When Piceatannol interfered with AML cells for 24 h,the ratio of autophagy marker LC3-I/LC3-I increased with the increase of concentration(r=0.672,r=0.549).When Piceatannol interfered with AML cells for48 h,the expression of Bcl-2 protein was down-regulated and caspase 3 was hydrolyzed and activated.At the same time,the activation level of Akt/NF-KB signaling pathway was inhibited to induce programmed death of AML cells.Piceatannol can also down-regulate the expression of MRPI and gradually weaken the chemotherapy resistance of AML drug-resistant cell lines,but it has a.weak effect on the expression of BCRP and almost no effect on MDRI.Conclusion:Piceatannol can inhibit the proliferation of AML cells and induce programmed death,which may be related to the inhibition of Akt/NF-KB signaling pathway,the hydrolysis of caspase-3 and the down-regulation of Bcl-2 protein expression,and the suppression of the expression of some drug resistance genes.