Binding of Released Bim to Mcl-1 is a Mechanism of Intrinsic Resistance to ABT-199 which can be Overcome by Combination with Daunorubicin or Cytarabine in AML Cells

被引:196
作者
Niu, Xiaojia [1 ,2 ]
Zhao, Jianyun [1 ]
Ma, Jun [1 ]
Xie, Chengzhi [3 ,4 ]
Edwards, Holly [3 ,4 ]
Wang, Guan [1 ]
Caldwell, J. Timothy [5 ,6 ]
Xiang, Shengyan [7 ]
Zhang, Xiaohong [7 ,8 ]
Chu, Roland [2 ,9 ]
Wang, Zhihong J. [2 ,9 ]
Lin, Hai [10 ]
Taub, Jeffrey W. [2 ,4 ,9 ]
Ge, Yubin [3 ,4 ]
机构
[1] Jilin Univ, Sch Life Sci, Natl Engn Lab AIDS Vaccine, Key Lab Mol Enzymol & Engn,Minist Educ, Changchun, Peoples R China
[2] Wayne State Univ, Sch Med, Dept Pediat, Detroit, MI 48201 USA
[3] Wayne State Univ, Sch Med, Dept Oncol, Detroit, MI USA
[4] Wayne State Univ, Sch Med, Barbara Ann Karmanos Canc Inst, Mol Therapeut Program, Detroit, MI USA
[5] Wayne State Univ, Sch Med, MD PhD Program, Detroit, MI USA
[6] Wayne State Univ, Sch Med, Canc Biol Grad Program, Detroit, MI USA
[7] USF Morsani, Coll Med, Dept Pathol & Cell Biol, Tampa, FL USA
[8] H Lee Moffitt Canc Ctr & Res Inst, Canc Biol & Evolut Program, Tampa, FL USA
[9] Childrens Hosp Michigan, Div Pediat Hematol Oncol, Detroit, MI 48201 USA
[10] Jilin Univ, Hosp 1, Dept Hematol & Oncol, Changchun, Peoples R China
关键词
ACUTE MYELOID-LEUKEMIA; DOWN-SYNDROME CHILDREN; BH3 MIMETIC ABT-199; CHEMOTHERAPY SENSITIVITY; ANTITUMOR-ACTIVITY; INHIBITOR ABT-199; BCL-2; INHIBITOR; VALPROIC ACID; FAMILY; EXPRESSION;
D O I
10.1158/1078-0432.CCR-15-3057
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Purpose: To investigate the molecular mechanism underlying intrinsic resistance to ABT-199. Experimental Design: Western blots and real-time RT-PCR were used to determine levels of Mcl-1 after ABT-199 treatment alone or in combination with cytarabine or daunorubicin. Immunoprecipitation of Bim and Mcl-1 were used to determine the effect of ABT-199 treatment on their interactions with Bcl-2 family members. Lentiviral short hairpin RNA knockdown of Bim and CRISPR knockdown of Mcl-1 were used to confirm their role in resistance to ABT-199. JC-1 assays and flow cytometry were used to determine drug-induced apoptosis. Results: Immunoprecipitation of Bim from ABT-199-treated cell lines and a primary patient sample demonstrated decreased association with Bcl-2, but increased association with Mcl-1 without corresponding change in mitochondrial outer membrane potential. ABT-199 treatment resulted in increased levels of Mcl-1 protein, unchanged or decreased Mcl-1 transcript levels, and increased Mcl-1 protein half-life, suggesting that the association with Bim plays a role in stabilizing Mcl-1 protein. Combining conventional chemotherapeutic agent cytarabine or daunorubicin with ABT-199 resulted in increased DNA damage along with decreased Mcl-1 protein levels, compared with ABT-199 alone, and synergistic induction of cell death in both AML cell lines and primary patient samples obtained from AML patients at diagnosis. Conclusions: Our results demonstrate that sequestration of Bim by Mcl-1 is a mechanism of intrinsic ABT-199 resistance and supports the clinical development of ABT-199 in combination with cytarabine or daunorubicin for the treatment of AML.
引用
收藏
页码:4440 / 4451
页数:12
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