α-Mangostin from Cratoxylum arborescens demonstrates apoptogenesis in MCF-7 with regulation of NF-κB and Hsp70 protein modulation in vitro, and tumor reduction in vivo
α-Mangostin from Cratoxylum arborescens demonstrates apoptogenesis in MCF-7 with regulation of NF-κB and Hsp70 protein modulation in vitro, and tumor reduction in vivo
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DOI：
10.2147/dddt.s66105
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发表时间：
2014-01-01
影响因子：
4.8
通讯作者：
Ali, Hapipah Mohd
中科院分区：
文献类型：
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作者：
Ibrahim, Mohamed Yousif;Hashim, Najihah Mohd;Ali, Hapipah Mohd
木立黄牛（Cratoxylum arborescens）是属于古提弗科（Guttiferae）的赤道植物。本研究分离出了α-山竹素(AM)并研究了其细胞死亡机制。 HCS用于检测核浓缩、线粒体膜电位、细胞通透性和细胞色素c的释放。使用荧光分析进行活性氧形成的研究。为了确定细胞死亡的机制，进行了人类凋亡蛋白质组分析仪测定。此外，还使用免疫荧光和免疫印迹检测了Bcl-2相关X蛋白（Bax）和B细胞淋巴瘤（Bcl）-2蛋白的水平。在治疗过程中评估了 3/7、8 和 9 等空间。使用 HCS 和蛋白质印迹，研究了核因子 kappa-B (NF-kappa B) 的贡献。 AM即使在30μg/mL的浓度下也显示出对癌细胞的选择性细胞毒性，而对正常细胞没有毒性，从而表明AM具有诱导肿瘤细胞死亡的特性。用AM处理MCF-7细胞通过细胞死亡转导信号促进细胞凋亡。这通过下调 Bcl-2 和上调 Bax 来调节线粒体膜电位，从而导致细胞色素 c 从线粒体释放到细胞质中。细胞色素 c 的释放激活了 caspace-9，而 caspace-9 又通过裂解的聚（ADP-核糖）聚合酶蛋白激活下游刽子手 caspace-3/7，从而导致细胞凋亡改变。 casspace 8 的增加表明外在途径的参与。这种类型的细胞凋亡被认为是通过外在和内在途径发生的，并防止 NF-κ B 从细胞质易位到细胞核。我们的结果表明，AM 通过 NF-kappa B、Bax/Bcl-2 和热休克蛋白 70 调节以及 casspace 的作用，促进 MCF-7 细胞凋亡。此外，在乳腺癌动物模型中，摄入 AM（30 和 60 mg/kg）可显着减小肿瘤大小。我们的结果表明 AM 是治疗乳腺癌的潜在有用药物。
Cratoxylum arborescens is an equatorial plant belonging to the family Guttiferae. In the current study, alpha-Mangostin (AM) was isolated and its cell death mechanism was studied. HCS was undertaken to detect the nuclear condensation, mitochondrial membrane potential, cell permeability, and the release of cytochrome c. An investigation for reactive oxygen species formation was conducted using fluorescent analysis. To determine the mechanism of cell death, human apoptosis proteome profiler assay was conducted. In addition, using immunofluorescence and immunoblotting, the levels of Bcl-2-associated X protein (Bax) and B-cell lymphoma (Bcl)-2 proteins were also tested. Caspaces such as 3/7, 8, and 9 were assessed during treatment. Using HCS and Western blot, the contribution of nuclear factor kappa-B (NF-kappa B) was investigated. AM had showed a selective cytotoxicity toward the cancer cells with no toxicity toward the normal cells even at 30 mu g/mL, thereby indicating that AM has the attributes to induce cell death in tumor cells. The treatment of MCF-7 cells with AM prompted apoptosis with cell death-transducing signals. This regulated the mitochondrial membrane potential by down-regulation of Bcl-2 and up-regulation of Bax, thereby causing the release of cytochrome c from the mitochondria into the cytosol. The liberation of cytochrome c activated caspace-9, which, in turn, activated the downstream executioner caspace-3/7 with the cleaved poly (ADP-ribose) polymerase protein, thereby leading to apoptotic alterations. Increase of caspace 8 had showed the involvement of an extrinsic pathway. This type of apoptosis was suggested to occur through both extrinsic and intrinsic pathways and prevention of translocation of NF-kappa B from the cytoplasm to the nucleus. Our results revealed AM prompt apoptosis of MCF-7 cells through NF-kappa B, Bax/Bcl-2 and heat shock protein 70 modulation with the contribution of caspaces. Moreover, ingestion of AM at (30 and 60 mg/kg) significantly reduced tumor size in an animal model of breast cancer. Our results suggest that AM is a potentially useful agent for the treatment of breast cancer.