Luciferase-based protein denaturation assay for quantification of radiofrequency field-induced targeted hyperthermia: developing an intracellular thermometer.
Luciferase-based protein denaturation assay for quantification of radiofrequency field-induced targeted hyperthermia: developing an intracellular thermometer.
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DOI：
10.3109/02656736.2012.666318
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Curley SA
中科院分区：
文献类型：
--
作者：
Raoof M;Zhu C;Kaluarachchi WD;Curley SA
几项研究已经报道了利用射频波远程激活纳米颗粒在细胞水平进行靶向热疗。到目前为止，量化细胞内热剂量的方法还没有报道。在这份报告中，我们研究了使用和不使用细胞内纳米颗粒时无线电波暴露和荧光素酶变性之间的关系。这些发现被用来设计一种策略，以量化原代人类肝癌细胞系中的靶向热剂量。热疗采用水浴或无创体外射频发生器(600W，13.56 MHz)。用生物发光法检测荧光素酶活性，Annexin V-FITC和碘化丙啶染色检测细胞活力。用Western-Blot分析热休克蛋白，观察到暴露在射频电场中的SNU449细胞在可测量的活力丧失之前发生了持续时间依赖的荧光素酶变性。在西妥昔单抗偶联金纳米粒(C225-AuNP)处理的细胞中，荧光素酶活性的损失更大。利用水浴实验的标准曲线，计算了细胞内的热剂量。经C225-AuNP处理的细胞在最初4分钟的RF暴露中积累的细胞内热剂量是未处理对照组的6.07倍。当癌细胞暴露在外部射频场中时，表现出剂量依赖性的蛋白质变性。荧光素酶变性试验可以用来定量RF暴露于癌细胞后的热剂量，无论有没有纳米颗粒。
Several studies have reported targeted hyperthermia at the cellular level using remote activation of nanoparticles by radiofrequency waves. To date, methods to quantify intracellular thermal dose have not been reported. In this report we study the relationship between radio wave exposure and luciferase denaturation with and without intracellular nanoparticles. The findings are used to devise a strategy to quantify targeted thermal dose in a primary human liver cancer cell line. Water-bath or non-invasive external RF generator (600W, 13.56 MHz) was used for hyperthermia exposures. Luciferase activity was measured using a bioluminescence assay and viability was assessed using Annexin V-FITC and Propidium iodide staining. Heat shock proteins were analyzed using western-blot analysis Duration-dependent luciferase denaturation was observed in SNU449 cells exposed to RF field that preceded measurable loss in viability. Loss of luciferase activity was higher in cetuximab-conjugated gold nanoparticle (C225-AuNP) treated cells. Using a standard curve from water-bath experiments, the intracellular thermal dose was calculated. Cells treated with C225-AuNP accumulated 6.07 times higher intracellular thermal dose than the untreated controls over initial 4 minutes of RF exposure. Cancer cells when exposed to an external RF field exhibit dose-dependent protein denaturation. Luciferase denaturation assay can be used to quantify thermal dose delivered after RF exposures to cancer cells with and without nanoparticles.
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DOI：
10.1023/b:jtan.0000033212.93809.e1
发表时间：
2004-01-01
影响因子：
4.4
作者：
Michnik, A;Klos, A;Drzazga, Z
通讯作者：
Drzazga, Z
DOI：
10.1158/1078-0432.ccr-10-2055
发表时间：
2010-12-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Glazer ES;Zhu C;Massey KL;Thompson CS;Kaluarachchi WD;Hamir AN;Curley SA
通讯作者：
Curley SA
影响因子：
15
作者：
Gota, Chie;Okabe, Kohki;Uchiyama, Seiichi
通讯作者：
Uchiyama, Seiichi
影响因子：
17.1
作者：
Creixell, Mar;Bohorquez, Ana C.;Rinaldi, Carlos
通讯作者：
Rinaldi, Carlos
影响因子：
14.8
作者：
Kumar, S.;Aaron, J.;Sokolov, K.
通讯作者：
Sokolov, K.