Effect of dual inhibition of apoptosis and autophagy in prostate cancer.
Effect of dual inhibition of apoptosis and autophagy in prostate cancer.
复制标题
DOI：
10.1002/pros.22487
复制
发表时间：
2012-09-01
期刊：
影响因子：
2.8
通讯作者：
DiPaola, Robert S.
中科院分区：
文献类型：
--
作者：
Saleem, Ahamed;Dvorzhinski, Dmitri;Santanam, Urmila;Mathew, Robin;Bray, Kevin;Stein, Mark;White, Eileen;DiPaola, Robert S.
现在，利用小分子BH3结构域模拟物，如ABT-737，靶向多种抗凋亡蛋白是可能的。鉴于最近的研究表明，自噬在抑制凋亡的背景下是一种对多种治疗药物的耐药机制，我们假设抑制自噬的抗疟疾药物羟基氯喹(HCQ)将增加ABT-737的细胞毒性。ABT-737和HCQ在体外对PC-3和LNCaP细胞的细胞毒性以及在异种移植小鼠模型中的细胞毒性进行了评估。自噬作为抗性机制的作用是通过siRNA敲除必要的自噬基因beclin1来评估的。用流式细胞仪检测ROS，用mCherry-Parkin报告评估有丝分裂。ABT-737诱导自噬是前列腺癌耐药的一种机制。HCQ抑制自噬的治疗作用增加了ABT-737的体内外细胞毒性。ABT-737通过免疫印迹和体内肿瘤免疫组织化学方法诱导LC-3和降低p62的表达。对ROS和线粒体的检测表明，ABT-737和HCQ产生ROS是细胞毒性的机制之一。我们证明，在体外和体内，HCQ抑制自噬增强了ABT-737的细胞毒性，LC-3和P62是未来临床试验中可评估的人体组织标志物，ROS诱导是细胞毒性的机制之一。这些结果支持在未来的临床研究中双靶向凋亡和自噬的新范式。
Targeting multiple anti-apoptotic proteins is now possible with the small molecule BH3 domain mimetics such as ABT-737. Given recent studies demonstrating that autophagy is a resistance mechanism to multiple therapeutic agents in the setting of apoptotic inhibition, we hypothesized that hydroxychloroquine (HCQ), an anti-malarial drug that inhibits autophagy, will increase cytotoxicity of ABT-737. Cytotoxicity of ABT-737 and HCQ was assessed in vitro in PC-3 and LNCaP cells, and in vivo in a xenograft mouse model. The role of autophagy as a resistance mechanism was assessed by siRNA knockdown of the essential autophagy gene beclin1. ROS was measured by flow cytometry, and mitophagy assessed by the mCherry-Parkin reporter. Induction of autophagy by ABT-737 was a mechanism of resistance in prostate cancer cell lines. Therapeutic inhibition of autophagy with HCQ increased cytotoxicity of ABT-737 both in vitro and in vivo. ABT-737 induced LC-3 and decreased p62 expression by immunoblot in cell lines and by immunohistochemistry in tumors in vivo. Assessment of ROS and mitochondria demonstrated that ROS production by ABT-737 and HCQ was a mechanism of cytotoxicity. We demonstrated that autophagy inhibition with HCQ enhances ABT-737 cytotoxicity in vitro and in vivo, that LC-3 and p62 represent assessable markers in human tissue for future clinical trials, and that ROS induction is a mechanism of cytotoxicity. These results support a new paradigm of dual targeting of apoptosis and autophagy in future clinical studies.
登录
查看更多内容
影响因子：
8
作者：
Ni Chonghaile T;Letai A
通讯作者：
Letai A
影响因子：
10.5
作者：
Guo, Jessie Yanxiang;Chen, Hsin-Yi;White, Eileen
通讯作者：
White, Eileen
DOI：
10.1073/pnas.0807694106
发表时间：
2009-02-24
影响因子：
11.1
作者：
Tal, Michal Caspi;Sasai, Miwa;Iwasaki, Akiko
通讯作者：
Iwasaki, Akiko
影响因子：
4.8
作者：
Castilla, Carolina;Congregado, Belen;Saez, Carmen
通讯作者：
Saez, Carmen
Bcl-2 同源结构域 3 模拟棉酚在癌细胞中诱导 Beclin 1 依赖性和 Beclin 1 独立的细胞保护性自噬
影响因子：
4.8
作者：
Gao, Ping;Bauvy, Chantal;Mehrpour, Maryam
通讯作者：
Mehrpour, Maryam