Inhibition of hexokinase-2 with targeted liposomal 3-bromopyruvate in an ovarian tumor spheroid model of aerobic glycolysis.
Inhibition of hexokinase-2 with targeted liposomal 3-bromopyruvate in an ovarian tumor spheroid model of aerobic glycolysis.
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DOI：
10.2147/ijn.s82818
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发表时间：
2015
影响因子：
8
通讯作者：
Amiji MM
中科院分区：
文献类型：
--
作者：
Gandham SK;Talekar M;Singh A;Amiji MM
本研究的目的是使用人卵巢腺癌细胞 (SKOV-3) 细胞的三维多细胞球体模型评估糖酵解标记物，特别是己糖激酶-2 (HK2) 的表达水平，并开发一种表皮生长因子受体靶向脂质体制剂，以改善 HK2 的抑制和 3-溴丙酮酸盐的细胞毒性 (3-BPA)。使用悬滴法开发多细胞 SKOV-3 肿瘤球体，并检查糖酵解标记物的表达水平。配制并表征了非靶向和表皮生长因子受体靶向的 3-BPA 脂质体制剂。使用共焦显微镜评估三维 SKOV-3 球体中脂质体制剂的渗透性和细胞摄取。通过细胞增殖和 HK2 酶测定，将 3-BPA 溶液形式的细胞毒性和 HK2 抑制潜力与相应的脂质体制剂进行比较。使用 96 孔悬滴法可重复地形成 SKOV-3 球体，第 5 天平均大小为 900 µm。发现低氧条件下的 HK2 酶活性水平高于常氧条件下（P<0.0001，学生 t 检验，未配对和双尾）。给药后 3、6 和 24 小时，50 µM 剂量的 3-BPA 脂质体制剂（非靶向和靶向）显示出比水溶液形式更有效的抑制作用（P<0.001，学生 t 检验，未配对和双尾）。同样，不同浓度（10 µM–100 µM）的 3-BPA 细胞毒活性表明，与 10 µM 和 25 µM 浓度的水溶液形式相比，脂质体制剂的细胞毒作用增强了 2–5 倍（P<0.0001，学生 t 检验，不配对和双尾）。通过悬滴法开发的 SKOV-3 球体可用作肿瘤有氧糖酵解模型，用于评估针对癌细胞糖酵解途径的治疗。将 3-BPA 封装在脂质体制剂中可改善多细胞球体模型中的渗透性、HK2 抑制和细胞毒性。
The objective of this study was to evaluate the expression levels of glycolytic markers, especially hexokinase-2 (HK2), using a three-dimensional multicellular spheroid model of human ovarian adenocarcinoma (SKOV-3) cells and to develop an epidermal growth factor receptor-targeted liposomal formulation for improving inhibition of HK2 and the cytotoxicity of 3-bromopyruvate (3-BPA). Multicellular SKOV-3 tumor spheroids were developed using the hanging drop method and expression levels of glycolytic markers were examined. Non-targeted and epidermal growth factor receptor-targeted liposomal formulations of 3-BPA were formulated and characterized. Permeability and cellular uptake of the liposomal formulations in three-dimensional SKOV-3 spheroids was evaluated using confocal microscopy. The cytotoxicity and HK2 inhibition potential of solution form of 3-BPA was compared to the corresponding liposomal formulation by using cell proliferation and HK2 enzymatic assays. SKOV-3 spheroids were reproducibly developed using the 96-well hanging drop method, with an average size of 900 µm by day 5. HK2 enzyme activity levels under hypoxic conditions were found to be higher than under normoxic conditions (P<0.0001, Student’s t-test, unpaired and two-tailed). Liposomal formulations (both non-targeted and targeted) of 3-BPA showed a more potent inhibitory effect (P<0.001, Student’s t-test, unpaired and two-tailed) at a dose of 50 µM than the aqueous solution form at 3, 6, and 24 hours post administration. Similarly, the cytotoxic activity 3-BPA at various concentrations (10 µM–100 µM) showed that the liposomal formulations had an enhanced cytotoxic effect of 2–5-fold (P<0.0001, Student’s t-test, unpaired and two-tailed) when compared to the aqueous solution form for both 10 µM and 25 µM concentrations. SKOV-3 spheroids developed by the hanging drop method can be used as a tumor aerobic glycolysis model for evaluation of therapies targeting the glycolytic pathway in cancer cells. Encapsulation of 3-BPA in a liposomal formulation improved permeability, HK2 inhibition, and cytotoxicity in the multicellular spheroid model.