IGF1-R抑制剂AEW541对伊马替尼杀伤SUP-B15细胞的增效作用  

Synergistic Effect of IGF1-R Inhibitor AEW541 on Imatinib Inducing SUP-B15 Cell Death

作　　者：王聪月 张雯雯 年丽 曹旭 席静静 郭纹彤 陈翀 WANG Cong-Yue;ZHANG Wen-Wen;NIAN Li;CAO Xu;XI Jing-Jing;GUO Wen-Tong;CHEN Chong(Blood Diseases Institute,Xuzhou Medical University,Department of Hematology,The Affiliated Hospital of Xuzhou Medical University;The First School of Clinical Medicine,Xuzhou Medical University,Xuzhou 221002,Jiangsu Province,China)

机构地区：[1]徐州医科大学血液病研究所,徐州医科大学附属医院血液科 [2]徐州医科大学第一临床医学院,江苏徐州221002

出　　处：《中国实验血液学杂志》2024年第6期1704-1710,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(82171762);江苏省普通高校研究生科研、实践创新计划项目(KYCX22-2904);江苏省高等学校大学生创新创业训练计划项目(202110313042Y)。

摘　　要：目的:探讨伊马替尼处理Ph^(+)ALL细胞株SUP-B15时是否出现细胞增殖受抑但不凋亡的耐受状态,探索IGF1-R抑制剂AEW541能否打破这种耐受状态以及相关机制。方法:使用不同浓度伊马替尼或AEW541处理SUP-B15细胞,Deep Blue试剂测定细胞增殖,Annexin V/7-AAD测定细胞凋亡;AEW541和伊马替尼联合处理SUP-B15细胞后流式细胞术测定细胞凋亡率,Western blot测定凋亡相关标志caspase-3和PARP1切割以及ABL下游信号STAT5、ERK1/2和AKT的磷酸化;STAT5和MEK-ERK1/2抑制剂进一步确认伊马替尼联合AEW541诱导SUP-B15细胞凋亡的关键机制。结果:伊马替尼单药处理能有效抑制SUP-B15细胞增殖,但没有显著增加SUP-B15细胞的凋亡率从而出现耐受状态;AEW541单药对SUP-B15细胞增殖与凋亡没有显著影响,但与伊马替尼合用时能显著增加SUP-B15细胞凋亡率及caspase-3和PARP1的切割;AEW541与伊马替尼联合处理SUP-B15较伊马替尼单药STAT5和ERK1/2磷酸化水平更低而对AKT磷酸化水平无明显影响;STAT5抑制剂AC-4-130而非MEK-ERK1/2抑制剂曲美替尼能诱导SUP-B15细胞凋亡。结论:SUP-B15细胞株可对伊马替尼呈现药物耐受,IGF1-R抑制剂AEW541可进一步降低STAT5的活化,继而增强伊马替尼对SUP-B15细胞的凋亡诱导作用,这为打破耐受提供新思路。Objective:To explore whether Ph^(+)acute lymphoblastic leukemia(ALL)cell line SUP-B15 treated with imatinib occurs a tolerant status charactered by cell proliferation suppression but apoptotic resistance,then evaluate whether IGF1-R inhibitor AEW541 can break this tolerance,and further explain its mechanisms.Methods:SUP-B15 cells were treated with different concentrations of imatinib or AEW541.Cell proliferation was assayed by Deep Blue,and apoptotic cells were determined by Annexin V/7-AAD staining.Apoptotic rate was measured by flow cytometry after co-treatment of imatinib and AEW541.Western blot was used to evaluate ABL downstream signals,including the phosphorylation of STAT5,ERK1/2,and AKT,as well as to detect cleaved caspase-3 and PARP1,the molecular signatures of apoptosis.Furthermore,an inhibitor of STAT5 or MEK-ERK1/2 was used to confirm the key mechanism of the combination of imatinib and AEW541 induced SUP-B15 cell apoptosis.Results:Imatinib monotherapy effectively suppressed the proliferation of SUP-B15 cells,but did not induce significant increase of apoptotic rate,leading to occurrence of tolerant status.AEW541 monotherapy did not dramatically affect the proliferation and apoptosis of SUP-B15 cells,but significantly increased apoptotic rate of SUP-B15 cells and cleavage of caspase-3 and PARP1 when combined with imatinib simultaneously.A combination of imatinib and AEW541 reduced STAT5 and ERK1/2 phosphorylation as compared with imatinib monotherapy in SUP-B15 cells,but had no impact on AKT phosphorylation.Apoptosis could be induced by STAT5 inhibitor AC-4-130,but not by MEK-ERK1/2 inhibitor trametinib in SUP-B15 cells.Conclusion:SUP-B15 cells treated with imatinib can establish drug tolerance.IGF1-R inhibitor AEW541 can further reduce STAT5 activation,thereby boosting the effect of apoptotic induction of imatinib on SUP-B15 cells.This research may provide a new idear to overcome imatinib tolerance.

关 键 词：药物耐受 细胞凋亡 Ph^(+)急性淋巴细胞白血病 伊马替尼 AEW541 

分 类 号：R733.71[医药卫生—肿瘤]