优化方法量化瘤治疗场 (TTFields) 诱导的质母细胞细胞膜的透性
Melisa Martinez-Paniagua1, Sabbir Khan1, Nikita W Henning1
1Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Unit 1002, BSRB S5.8116b, Houston, TX 77030, USA.
Methods and protocols
|February 25, 2025
概括
瘤治疗场 (TTFields) 透人类质母细胞细胞,这一发现得到了两种新方法的验证. 这些技术量化了TTFields的数量.
科学领域:
- 在瘤学瘤学.
- 生物物理学的生物物理.
- 生物化学 生物化学
背景情况:
- 质母细胞瘤 (GBM) 是一种具有不良预后的侵袭性脑癌.
- 瘤治疗场 (TTFields) 在改善GBM患者的存活率方面表现有前途.
- 以前评估TTFields对GBM细胞的影响的方法在区分活细胞与死细胞吸收方面存在局限性.
研究的目的:
- 优化和验证两种独立的方法来量化 TTFields 诱导的人类 GBM 细胞膜透.
- 建立可靠的测试来评估TTFields对GBM细胞膜的影响.
主要方法:
- 流细胞测量被优化以测量TTFields暴露后的GBM细胞中的FITC-dextran探针 (4-20kDa) 的平均光强度.
- 测量乳酸脱酶 (LDH) 释放并将其正常化为细胞活力 (CellTiter-Glo测定),以确定LDH/CTG比率.
- 透效应与Triton-X-100进行了比较,并与卡博普拉丁治疗结合评估.
主要成果:
- 流细胞测量显示,在TTFields治疗的GBM细胞中,FITC-dextran的摄取量在4kDa (p=0.016) 和20kDa (p=0.031) 探针中显著增加.
- 与对照细胞 (1.08 ± 0.08,p < 0.0001) 相比,在TTFields暴露的细胞中,LDH/CTG比率显著更高 (1.47 ± 0.15).
- 这两种方法都可重复地证明了TTFields诱导的细胞膜透性,并且与碳柏金结合时有效.
结论:
- 两种经过验证的,具有成本效益的方法 (流细胞计和LDH/CTG比) 可以可靠地量化TTFields诱导的GBM细胞膜透.
- 这些方法为时间依赖性评估和评估TTFields与其他疗法结合的效果提供了一个强大的平台.
- 这些发现有助于更好地了解TTFields在GBM治疗中的作用机制.
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