Induction of Apoptosis by Functionalized Fullerene-based Sonodynamic Therapy in HL-60 cells.
Induction of Apoptosis by Functionalized Fullerene-based Sonodynamic Therapy in HL-60 cells.
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发表时间：
2016-06
影响因子：
2
通讯作者：
N. Yumita;Takahiro Watanabe;Fu-shih Chen;Y. Momose;S. Umemura
中科院分区：
文献类型：
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作者：
N. Yumita;Takahiro Watanabe;Fu-shih Chen;Y. Momose;S. Umemura
超声波由于其穿透深层组织的能力而具有较少的能量衰减和最小的不良副作用，已被广泛用于医学诊断和治疗。功能化富勒烯，如多羟基富勒烯（PHF），由于其水溶性和作为碳纳米材料在肿瘤成像和治疗中的潜在应用而引起了特别的关注。本研究探讨声动力学诱导的细胞凋亡使用PHF。在PHF存在下用2 MHz连续超声处理细胞悬液3 min，并通过使用共聚焦显微镜、DNA片段化（琼脂糖凝胶电泳后的梯形图）和caspase-3活化的细胞形态来评估细胞凋亡。在以下条件下从培养皿底部对细胞进行超声辐照：频率，2 MHz;输出功率，3 W/cm（2）电子自旋共振用于测量活性氧。声动力学暴露（超声和PHF）后的凋亡细胞的数量显着高于其他治疗，如超声单独和PHF单独产生。此外，在声动力学处理的细胞中观察到DNA片段化、半胱天冬酶-3活化和增强的2，2，6，6-四甲基-4-哌啶基氧基（4 oxoTEMPO）形成。组氨酸，一个众所周知的活性氧清除剂，显着抑制声动力学诱导的细胞凋亡，caspase-3激活和4 oxoTEMPO形成。PHF声动力作用诱导HL-60细胞发生凋亡，其特征为细胞皱缩、凋亡小体碎裂等一系列典型的形态学特征。声动力诱导的细胞凋亡，半胱天冬酶-3激活，和4 oxoTEMPO形成由于组氨酸和色氨酸的显着抑制表明，活性氧物种，如单线态氧，参与声动力诱导的细胞凋亡。这些结果表明，PHF介导的声动力治疗可以触发caspase依赖的细胞凋亡和氧化损伤，从而可能在凋亡信号级联中发挥重要作用。
Ultrasound has been widely utilized for medical diagnosis and therapy due to its ability to penetrate deep-seated tissue with less attenuation of energy and minimal undesirable side-effects. Functionalized fullerenes, such as polyhydroxy fullerene (PHF), have attracted particular attention due to their water solubility and potential application in tumor imaging and therapy as carbon nanomaterials. The present study investigated sonodynamically-induced apoptosis using PHF. Cell suspensions were treated with 2-MHz continuous ultrasound in the presence of PHF for 3 min and apoptosis was assessed by cell morphology using confocal microscopy, fragmentation of DNA (ladder pattern after agarose-gel electrophoresis) and caspase-3 activation. Cells were ultrasound-irradiated from the bottom of the culture dishes under the following condition: frequency, 2 MHz; output power, 3 W/cm(2) Electron spin resonance was used to measure reactive oxygen species. The number of apoptotic cells after sonodynamic exposure (ultrasound and PHF) was significantly higher than produced from other treatments, such as ultrasound alone and PHF alone. Furthermore, DNA fragmentation, caspase-3 activation and enhanced 2,2,6,6-tetramethyl-4-piperidinyloxy (4oxoTEMPO) formation were observed in the sonodynamically-treated cells. Histidine, a well-known reactive oxygen scavenger, significantly inhibited sonodynamically-induced apoptosis, caspase-3 activation and 4oxoTEMPO formation. Sonodynamic therapy with PHF induced apoptosis that was characterized by a series of typical morphological features, such as shrinkage of the cell and fragmentation into membrane-bound apoptotic bodies, in HL-60 cells. The significant inhibition of sonodynamically-induced apoptosis, caspase-3 activation, and 4oxoTEMPO formation due to histidine and tryptophan suggests that reactive oxygen species, such as singlet oxygen, are involved in the sonodynamic induction of apoptosis. These findings indicate that PHF-mediated sonodynamic therapy can trigger caspase-dependent apoptosis and oxidative injury, thus possibly playing a vital role in apoptotic signaling cascades.