SNS-032诱导弥漫性大B淋巴瘤OCI-LY-19细胞株凋亡及其作用机制的研究

摘要

AIM:To investigate the effects of SNS-032 on the apoptosis of human diffuse large B-cell lymphoma cell line OCI-LY-19 and its underlying mechanisms.METHODS:OCI-LY-19 cells were treated with different concentrations of SNS-032.The cell viability was measured by MTS assay.The cell death was analyzed by propidium iodide (PI) staining,and the cel apoptosis was analyzed by flow cytometry with Annexin V/PI staining.The expression of apoptosis-re-lated and prliferation-related proteins was analyzed by Western blotting.RESULTS:The viability and proliferation of OCI-LY-19 cells was inhibited by SNS-032 and the IC50 was 0.358 μmol/L.The percentage of apoptotie cells was increased by SNS-032 in a dose-and time-dependent manner.Poly(ADP-ribose) polymerase (PARP) protein was cleaved after SNS-032 treatment.The expression of apoptosis-related proteins,procaspase-3,procaspase-9,X-linked inhibitor of apoptosis protein (XIAP) and myeloid cell leukemia-1 (Mcl-1),and proliferation-related proteins,protein serine/threonine kinase (Akt),phosphory-lated Akt (p-Akt),signal transductor and activator of transcription 5 (STAT5),phosphorylted STAT5 (p-STAT5) and phosphorylated extracellular signal-regulated kinase (p-ERK),was down-regulated by SNS-032 in a dose-and time-dependent manner.Cleaved caspase-3 and cleaved caspase-9 expression levels were devated after SNS-032 treatment.The expression of total ERK was not obviously changed.CONCLUSION:SNS-032 can induce apoptosis of OCI-LY-19 cells and the mechanism may associate with the cleavage of PARP,the down-regulation of apoptosis-related proteins and the inhibition of proliferation-related JAK/STAT,MEK/ERK and PI3K/Akt signaling pathways.%目的:研究SNS-032对弥漫性大B淋巴瘤细胞株OCI-LY-19凋亡的影响并探讨其作用机制.方法:应用MTS法观察不同浓度的SNS-032对OCI-LY-19细胞活力的影响；用PI单染法观察SNS-032对OCI-LY-19细胞诱导死亡的影响；Annexin V/PI双染流式细胞术分析不同浓度SNS-032对OCI-LY-19细胞凋亡的影响；Western blotting检测细胞凋亡相关蛋白和细胞增殖相关蛋白的表达情况.结果:SNS-032明显抑制OCI-LY-19细胞活力,半数抑制浓度为0.358 μmol/L；随SNS-032作用时间延长与浓度升高,凋亡细胞数逐渐增多；SNS-032可以时间与剂量依赖性地引起凋亡相关蛋白聚腺苷二磷酸-核糖聚合酶(PARP)的切割,caspase-3前体(procaspase-3)、caspase-9前体(procaspase-9)、X连锁凋亡抑制蛋白(XIAP)与髓样细胞白血病1(Mcl-1)的表达下降,而cleaved caspase-3和cleaved caspase-9表达明显增加；与细胞增殖相关的蛋白丝／苏氨酸激酶(Akt)、磷酸化Akt(p-Akt)、信号转导子及转录激活子5(STAT5)、磷酸化STAT5(p-STAT5)、细胞外信号调节激酶(ERK)和磷酸化ERK(p-ERK)表达下降,而ERK总蛋白无明显变化.结论:SNS-032能抑制弥漫性大B淋巴瘤细胞株OCI-LY-19的生长,诱导其凋亡,可能的机制是抑制了相关抗凋亡蛋白的表达,激活了caspase级联反应,同时抑制了与细胞增殖相关的JAKs/STATs、MEK/ERK和PI3K-Akt信号转导通路的表达与活化.