Suppression of NF-κB survival signaling by nitrosylcobalamin sensitizes neoplasms to the anti-tumor effects of Apo2L/TRAIL (Retracted Article)
Suppression of NF-κB survival signaling by nitrosylcobalamin sensitizes neoplasms to the anti-tumor effects of Apo2L/TRAIL (Retracted Article)
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DOI：
10.1074/jbc.m306111200
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发表时间：
2003-10-10
影响因子：
4.8
通讯作者：
Lindner, DJ
中科院分区：
文献类型：
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作者：
Chawla-Sarkar, M;Bauer, JA;Lindner, DJ
我们以前已经证明了亚硝基钴胺素（NO-Cbl）的抗肿瘤活性，维生素B12的类似物，提供一氧化氮（NO），并增加肿瘤坏死因子相关凋亡诱导配体（Apo 2L/TRAIL）及其受体在人类肿瘤中的表达。本研究的具体目的是检查NO-Cbl是否可以使耐药黑色素瘤对Apo 2L/TRAIL敏感。在恶性黑色素瘤和非致瘤性黑素细胞和成纤维细胞系中评估NO-Cbl和Apo 2L/TRAIL的抗增殖作用。用NO-Cbl和Apo 2L/TRAIL处理携带人黑素瘤A375异种移植物的无胸腺裸鼠。通过TUNEL法测定细胞凋亡，并通过检查细胞凋亡关键介质的水平和活性来证实。通过测定DNA结合、荧光素酶报告活性、IkappaB α的磷酸化状态和体外IKK活性来确定NF-κ B的活化状态。NO-Cbl使Apo 2L/TRAIL抗性黑色素瘤细胞系对Apo 2L/TRAIL的生长抑制敏感，但对正常细胞系的影响最小。NO-Cbl和Apo 2L/TRAIL对A375异种移植物具有协同抗肿瘤活性。在Apo 2L/TRAIL耐药肿瘤细胞中，先用NO-Cbl再用Apo 2L/TRAIL处理诱导凋亡，其特征在于半胱天冬酶-3、半胱天冬酶-8和PARP的裂解。NO-Cbl抑制IKK活化，其特征在于IkappaB α磷酸化降低和NF-κ B DNA结合活性抑制。NO-Cbl抑制Apo 2L/TRAIL和TNF-α介导的转染NF-κ B驱动的荧光素酶报告基因的激活。XIAP是一种凋亡抑制剂，可被NO-Cbl灭活。NO-Cbl处理使得Apo 2L/TRAIL抗性恶性肿瘤在体外和体内对Apo 2L/TRAIL的抗肿瘤作用敏感。NO-Cbl和Apo 2L/TRAIL的使用利用了两种试剂的肿瘤特异性性质，代表了一种有前途的抗癌组合。
We have previously demonstrated the anti-tumor activity of nitrosylcobalamin (NO-Cbl), an analog of vitamin B12 that delivers nitric oxide (NO) and increases the expression of tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) and its receptors in human tumors. The specific aim of this study was to examine whether NO-Cbl could sensitize drug-resistant melanomas to Apo2L/TRAIL. Antiproliferative effects of NO-Cbl and Apo2L/TRAIL were assessed in malignant melanomas and non-tumorigenic melanocyte and fibroblast cell lines. Athymic nude mice bearing human melanoma A375 xenografts were treated with NO-Cbl and Apo2L/TRAIL. Apoptosis was measured by TUNEL and confirmed by examining levels and activity of key mediators of apoptosis. The activation status of NF-kappaB was established by assaying DNA binding, luciferase reporter activity, the phosphorylation status of IkappaBalpha, and in vitro IKK activity. NO-Cbl sensitized Apo2L/TRAIL-resistant melanoma cell lines to growth inhibition by Apo2L/TRAIL but had minimal effect on normal cell lines. NO-Cbl and Apo2L/TRAIL exerted synergistic antitumor activity against A375 xenografts. Treatment with NO-Cbl followed by Apo2L/TRAIL induced apoptosis in Apo2L/TRAIL-resistant tumor cells, characterized by cleavage of caspase-3, caspase-8, and PARP. NO-Cbl inhibited IKK activation, characterized by decreased phosphorylation of IkappaBalpha and inhibition of NF-kappaB DNA binding activity. NO-Cbl suppressed Apo2L/TRAIL- and TNF-alpha-mediated activation of a transfected NF-kappaB-driven luciferase reporter. XIAP, an inhibitor of apoptosis, was inactivated by NO-Cbl. NO-Cbl treatment rendered Apo2L/TRAIL- resistant malignancies sensitive to the anti-tumor effects of Apo2L/TRAIL in vitro and in vivo. The use of NO-Cbl and Apo2L/TRAIL capitalizes on the tumor-specific properties of both agents and represents a promising anti-cancer combination.