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  

Mechanism of SNS-032-induced apoptosis of human diffuse large B-cell lymphoma cell line OCI-LY-19

作　　者：师宪平[1,2] 蓝晓莹[1] 温创宇[3,4] 陈鑫[2] 刘焕亮[3,4] 黄美近[4] 

机构地区：[1]广州医科大学生物化学教研室,广东广州510182 [2]广州医科大学免疫研究所,广东广州510182 [3]中山大学胃肠病学研究所,广东广州510655 [4]中山大学附属第六医院,广东广州510655

出　　处：《中国病理生理杂志》2013年第7期1186-1191,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81100378);广州市科信局应用基础研究专项重点项目(No.2012J4100014);广东省医学科研基金资助项目(No.B2012159)

摘　　要：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0.358μmol/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cleavedcaspase-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。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 cell apoptosis was analyzed by flow cytometry with Annexin V / PI staining.The expression of apoptosis-related and proliferation-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 apoptotic 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）,phosphorylated 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 elevated 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.

关 键 词：SNS-032 弥漫性大B淋巴瘤 细胞凋亡 

分 类 号：R733.4[医药卫生—肿瘤]