PI3Kδ抑制剂CAL-101对淋巴瘤细胞系Raji和SUDHL-10的作用及其机制研究  

Inhibitory effects of the phosphoinostitide-3'-kinase delta inhibitor CAL-101 on Raji and SUDHL-10 lymphoma cells and its relative mechanism

作　　者：王亚非[1] 夏冰[1] 屈福莲[1] 李晓武[1] 郭姗琦[1] 袁田[1] 赵伟鹏[1] 张翼鷟[1] 

机构地区：[1]天津医科大学肿瘤医院血液科,国家肿瘤临床医学研究中心,天津市肿瘤防治重点实验室,天津市300060

出　　处：《中国肿瘤临床》2015年第3期135-140,共6页Chinese Journal of Clinical Oncology

基　　金：天津市教委创新人才中青年骨干培养计划基金项目资助~~

摘　　要：目的:探讨PI3Kδ抑制剂CAL-101对弥漫大B细胞淋巴瘤细胞系SUDHL-10和Burkitt淋巴瘤细胞系Raji的作用及其相关机制,为这类疾病的治疗提供新的思路。方法:用不同浓度的CAL-101处理Burkitt淋巴瘤细胞系Raji和弥漫大B细胞淋巴瘤细胞系SUDHL-10,以MTT法检测CAL-101对两种细胞系的增殖抑制作用;以Annexin V/PI流式细胞术和DAPI染色法检测细胞的凋亡情况;迁移实验检测淋巴瘤细胞向淋巴瘤基质细胞系HK的迁移比例;Western blot法检测CAL-101处理后淋巴瘤细胞表达磷酸化ERK的变化;MTT法结合Calcu Syn software软件分析检测CAL-101是否可协同硼替佐米抑制淋巴瘤细胞的增殖。结果:5μmol/L及更高浓度的CAL-101对Raji细胞和SUDHL-10细胞的增殖有明显的抑制作用,且呈剂量依赖性。5、10、15、20μmol/L CAL-101作用于Raji细胞48 h,细胞增殖抑制率分别为(29.17±1.23)%、(38.15±1.51)%、(46.46±1.78)%、(55.8±2.01)%,空白对照组为(1.15±0.02)%(P<0.05)。作用于SUDHL-10 48 h,细胞增殖抑制率也逐渐增加(P<0.05)。CAL-101可诱导淋巴瘤细胞的凋亡,10、20μmol/L CAL-101作用于Raji细胞24 h,Annexin V-FITC/PI双标法显示其凋亡率分别为(22.69±3.83)%和(49.96±7.36)%,均高于对照组(5.23±2.04)%(P<0.05);作用于SUDHL-10细胞同样得到相似的结果(P<0.05)。CAL-101可显著降低淋巴瘤细胞向基质细胞的迁移,且对Raji细胞和SUDHL-10细胞的迁移率的抑制作用也呈浓度依赖性,差异均具有统计学意义(P<0.05);Western blot检测发现CAL-101处理细胞后磷酸化ERK的表达明显降低;CAL-101与硼替佐具有协同作用,可显著抑制淋巴瘤细胞的增殖,CI<1。结论:PI3Kδ抑制剂CAL-101可抑制淋巴瘤细胞Raji和SUDHL-10的增殖,诱导凋亡,抑制其向淋巴瘤基质细胞的迁移,其机制可能通过阻断ERK信号途径而实现;CAL-101有望为侵袭性淋巴瘤的治疗带来希望。Objective：To detect the inhibitory effects of CAL-101, a selective inhibitor of phosphoinostitide-3＇-kinase delta （PI3Kδ）, on Burkitt＇s lymphoma cell line Raji and diffused large B-cell lymphoma cell line SUDHL-10 and elucidate its relative mechanism. Methods：Raji and SUDHL-10 cells were treated with various concentrations of CAL-101. Methyl thiazolyl tetrazolium （MTT） assay was performed to determine the inhibitory effect of CAL-101 on lymphoma cells, and cell apoptosis was measured by Annexin V/PI and DAPI staining. Migration assays were performed with transwell to detect the migration of lymphoma cells derived from the stromal cell line HK. Western blot was used to detect the phosphorylation status of the ERK pathway. MTT and CalcuSyn software analyses were preformed to detect whether or not combining CAL-101 with bortezomib induces synergistic cytoxicity. Results：CAL-101 at con-centrations of 5, 10, 15, and 20μmol/L inhibited cell proliferation in a dose-dependent manner. The proliferation rates of the Raji cells treated with 5, 10, 15, and 20μmol/L for 48 h were 29.17%± 1.23%, 38.15%± 1.51%, 46.46%± 1.78%, and 55.8%± 2.01%, respec-tively, which were significantly higher （P〈0.05） than that of the control group （1.15% ± 0.02%）. Similar results were found in the SUDHL-10 cells after treatment with CAL-101 （P〈0.05）. CAL-101 also exerted an apoptotic effect on the lymphoma cells. The apop-totic rates of the Raji cells treated with CAL-101 for 21 h were 22.69%± 3.83%and 49.96%± 7.36%, respectively, which were signifi-cantly higher （P〈0.05） than that of the control group （5.23%± 2.04%）. Similar results were found in the SUDHL-10 cells （P〈0.05）.＆amp;nbsp;Treatment with 5 and 10 μmol/L CAL-101 dose-dependently inhibited the migration activity of lymphoma cells to stromal cells （P〈0.05）. Western blot analysis showed that the expression level of ERK phosphorylation protein was significantly downregulated in the cells treated with CAL-101. A synergist

关 键 词：淋巴瘤 RAJI细胞 SUHDL-10细胞 PI3Kδ CAL-101 ERK通路 

分 类 号：R733.1[医药卫生—肿瘤]