Bcl-2 and Mn-SOD antisense oligodeoxynucleotides and a glutamine-enriched diet facilitate elimination of highly resistant B16 melanoma cells by tumor necrosis factor-α and chemotherapy
Bcl-2 and Mn-SOD antisense oligodeoxynucleotides and a glutamine-enriched diet facilitate elimination of highly resistant B16 melanoma cells by tumor necrosis factor-α and chemotherapy
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DOI：
10.1074/jbc.m507471200
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发表时间：
2006-01-06
影响因子：
4.8
通讯作者：
Estrela, JM
中科院分区：
文献类型：
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作者：
Benlloch, M;Mena, S;Estrela, JM
（1）线粒体谷胱甘肽（mtGSH）耗竭增加Bcl- 2过表达的B16黑色素瘤（B16 M）- F10细胞（高转移潜能）对肿瘤坏死因子-α（TNF-α）诱导的氧化应激和死亡的敏感性。在体内，通过给小鼠（其中B16 M-F10在足垫中作为实体瘤生长）喂食富含L-谷氨酰胺（L-Gln）的饮食，实现B16 M-F10细胞中的mtGSH耗竭，这促进了肿瘤细胞中谷氨酰胺酶活性的增加、细胞溶质L-谷氨酸的积累以及GSH转运到线粒体中的竞争性抑制。将使用抗Met- 72单克隆抗体和流式细胞术偶联的细胞分选分离的L-Gln适应的B16 M-F10细胞注射到门静脉中以产生肝转移。在通过相同方法从肝脏分离并用TNF-α和反义Bcl- 2寡脱氧核苷酸处理的L-Gln适应的侵袭性（iB 16 M-Gln（+））细胞中，存活率降低至接近12%. iB 16 M-Gln（+）细胞死亡与O-2生成增加相关（中心点）。和H2 O2，线粒体通透性转换孔复合物的开放，以及促凋亡分子信号的释放。细胞死亡机制的激活被GSH酯诱导的mtGSH补充所阻止。抗氧化应激的幸存者表现出适应性反应，包括过表达含锰超氧化物歧化酶（Mn-SOD）和过氧化氢酶活性。通过用双重抗-反义疗法（Bcl- 2和SOD 2反义寡脱氧核苷酸）和TNF-α处理iB 16 M-Gln（+）细胞，转移性细胞存活率降低至接近1%.化疗（紫杉醇加柔红霉素）很容易消除这一最低比例的幸存者。这一贡献确定了关键分子，可以依次靶向，以促进消除高耐药转移细胞。
(1)Mitochondrial glutathione ( mtGSH) depletion increases sensitivity of Bcl- 2- overexpressing B16 melanoma ( B16M)- F10 cells ( high metastatic potential) to tumor necrosis factor- alpha( TNF- alpha)- induced oxidative stress and death in vitro. In vivo, mtGSH depletion in B16M- F10 cells was achieved by feeding mice ( where the B16M-F10 grew as a solid tumor in the footpad) with an L- glutamine ( L- Gln)- enriched diet, which promoted in the tumor cells an increase in glutaminase activity, accumulation of cytosolic L- glutamate, and competitive inhibition of GSH transport into mitochondria. L- Gln- adapted B16M- F10 cells, isolated using anti- Met- 72 monoclonal antibodies and flow cytometry- coupled cell sorting, were injected into the portal vein to produce hepatic metastases. In L- Gln- adapted invasive ( iB16M- Gln(+)) cells, isolated from the liver by the same methodology and treated with TNF- alpha and an antisense Bcl- 2 oligodeoxynucleotide, viability decreased to similar to 12%. iB16M-Gln (+) cell death associated with increased generation of O-2(center dot). and H2O2, opening of the mitochondrial permeability transition pore complex, and release of proapoptotic molecular signals. Activation of cell death mechanisms was prevented by GSH ester- induced mtGSH replenishment. The oxidative stress- resistant survivors showed an adaptive response that includes overexpression of manganese-containing superoxide dismutase ( Mn- SOD) and catalase activities. By treating iB16M- Gln (+) cells with a double anti- antisense therapy ( Bcl- 2 and SOD2 antisense oligodeoxynucleotides) and TNF- alpha, metastatic cell survival decreased to similar to 1%. Chemotherapy ( taxol plus daunorubicin) easily removed this minimum percentage of survivors. This contribution identifies critical molecules that can be sequentially targeted to facilitate elimination of highly resistant metastatic cells.