Enhanced Cytotoxic Effect of Ara-C by Low Energy Ultrasound to HL-60 Cells

低能超声增强 Ara-C 对 HL-60 细胞的细胞毒作用

• 批准号: 09671777
• 负责人: TSUJITA Naotaka
• 金额: $ 1.15万
• 依托单位: Fukuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671777/
• 关键词: Ultrasound; Chemotherapy; Lymphocytes; Ara-C; SEM

Clonogenic assay was tested in order to determine the effects of low energy ultrasound (US) on HL-60 cells in the presence of cytosine arabinoside (Ara-C). HL-60 cells were exposed to ultrasound at an intensity of 0.3 W/cmィイD12ィエD1 (48 kHz). Cells were then cultured by methylcellulose method for 8 days and the number of colonies was statistically analyzed (one-way and two-way ANOVA). Ultrasound exposure alone for 120 seconds resulted in no significant decrease of colonies compared to non-treated cells (PィイD20ィエD2 = 0.1426). Significant differences (PィイD20ィエD2 < 0.005) were obtained between ultrasound treated and untreated cells in the presence of various concentrations of Ara-C (2×10ィイD1-9ィエD1, 1×10ィイD1-8ィエD1, 2×10ィイD1-8ィエD1, 5×10ィイD1-8ィエD1, 1×10ィイD1-7ィエD1M). Dose response studies revealed a significant enhancement of the anti-proliferative effects on colonies of HL-60 cells treated with ultrasound and Ara-C (two-way ANOVA/US ; PィイD20ィエD2=0.0000, 8.16%, Ara-C ; PィイD20ィエD2=0.0000, 88.91%). At lower concentration of Ara-C (0, 2×10ィイD1-9ィエD1, 1×10ィイD1-8ィエD1, 2×10ィイD1-8ィエD1M), the effects of ultrasound were greater than that of Ara-C (two-way ANOVA/US ; PィイD20ィエD2=0.0000, 71.33%, Ara-C ; PィイD20ィエD2=0.0000, 14.99%). It is suggested that ultrasound irradiation is more effective at relatively low non-toxic Ara-C concentration levels. Morphological evaluation of ultrasound irradiated cells with scanning electron microscopy showed minor disruption of cell surface and disappearance of microvilli. These observations suggests that low energy ultrasound altered the cell membrane thus resulting in change in Ara-C uptake into HL-60 cells.

采用克隆形成实验研究了低能量超声对HL-60细胞在阿糖胞苷（Ara-C）存在下的作用。HL-60细胞暴露于强度为0.3 W/cm × 12 kHz × 12 D1（48 kHz）的超声。然后通过甲基纤维素法培养细胞8天，并对集落数进行统计学分析（单因素和双因素ANOVA）。与未处理的细胞相比，单独超声暴露120秒未导致集落的显著减少（P <0.05）。在不同浓度的Ara-C（2×10 μ g/ml D1-9 μ g/ml D1，1×10 μ g/ml D1-8 μ g/ml D1，2×10 μ g/ml D1-8 μ g/ml D1，5×10 μ g/ml D1 - 8 μ g/ml D1，1 × 10 μ g/ml D1-7 μ g/ml D1 M）存在下，超声处理的细胞与未处理的细胞相比，差异有显著性（P < 0.005）。剂量反应研究显示，超声和Ara-C处理的HL-60细胞集落的抗增殖作用显著增强（双因素ANOVA/US ; P <0.0000，8.16%，Ara-C ; P <0.0000，88.91%）。在低浓度Ara-C（0、2×10 μ g/L D1-9 μ g/L D1、1×10 μ g/L D1 - 8 μ g/L D1 M）时，超声作用大于Ara-C（双因素方差分析/US ; P < 0.0000，71.33%，Ara-C ; P < 0.0000，14.99%）。有人建议，超声辐照是更有效的，在相对较低的无毒Ara-C浓度水平。超声辐照细胞的形态学评价与扫描电子显微镜显示轻微的破坏细胞表面和消失的微绒毛。这些观察结果表明，低能量超声改变了细胞膜，从而导致HL-60细胞的Ara-C摄取的变化。