Real-time monitoring of cisplatin cytotoxicity on three-dimensional spheroid tumor cells.
Real-time monitoring of cisplatin cytotoxicity on three-dimensional spheroid tumor cells.
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DOI：
10.2147/dddt.s108004
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
An SS
中科院分区：
文献类型：
--
作者：
Baek N;Seo OW;Lee J;Hulme J;An SS
三维（3D）细胞培养是一项强大的技术，用于监测和理解发育中的癌症和神经生物学、组织工程和药物开发中的各种细胞机制。与二维（2D）培养相比，3D系统可以更好地与体内模型相关。几个因素，如细胞类型、存活率、增殖率、基因和蛋白质表达模式，决定了一个特定的细胞系是否可以适应3D系统。3D系统可以克服2D培养在细胞间通信和细胞网络方面的一些局限性，这对于理解分化、结构组织、形状以及与其他细胞或器官的扩展连接至关重要。在这里，抗癌药物顺铂，也被称为顺式二胺二氯铂（II）或CDDP，对三磷酸腺苷（ATP）生成的影响进行了研究，使用三维球体形成细胞和实时监测7天。首先，筛选12个细胞系形成三维球体的能力：前列腺（DU145）、睾丸（F9）、胚胎成纤维细胞（NIH-3T3）、肌肉（C2C12）、胚胎肾（293T）、神经母细胞瘤（SH-SY5Y）、腺癌肺泡基底上皮细胞（A549）、宫颈癌（HeLa）、HeLa污染物（HEp2）、垂体上皮样细胞（GH3）、胚胎细胞（PA317）和骨肉瘤（U-2OS）细胞。其中选择NIH-3T3、C2C12、293T、SH-SY5Y、A549、HeLa、PA317、u - 2s 8个细胞系；实时监测5种CDDP细胞毒性：HeLa、A549、293T、SH-SY5Y和U-2OS。添加50 μM CDDP后1天ATP的生成被阻断，但仅在处理后4天即可观察到对HeLa、A549、SH-SY5Y和u - 2s细胞的细胞毒性。在293T细胞中，CDDP未能完全杀死培养物，1天后ATP的生成仅被部分阻断。这表明293T细胞可能对CDDP耐药或药物代谢清除。实时监测和ATP测量直接证实了CDDP的细胞毒性，表明CDDP可能干扰线粒体活性。
Three-dimensional (3D) cell cultivation is a powerful technique for monitoring and understanding diverse cellular mechanisms in developmental cancer and neuronal biology, tissue engineering, and drug development. 3D systems could relate better to in vivo models than two-dimensional (2D) cultures. Several factors, such as cell type, survival rate, proliferation rate, and gene and protein expression patterns, determine whether a particular cell line can be adapted to a 3D system. The 3D system may overcome some of the limitations of 2D cultures in terms of cell–cell communication and cell networks, which are essential for understanding differentiation, structural organization, shape, and extended connections with other cells or organs. Here, the effect of the anticancer drug cisplatin, also known as cis-diamminedichloroplatinum (II) or CDDP, on adenosine triphosphate (ATP) generation was investigated using 3D spheroid-forming cells and real-time monitoring for 7 days. First, 12 cell lines were screened for their ability to form 3D spheroids: prostate (DU145), testis (F9), embryonic fibroblast (NIH-3T3), muscle (C2C12), embryonic kidney (293T), neuroblastoma (SH-SY5Y), adenocarcinomic alveolar basal epithelial cell (A549), cervical cancer (HeLa), HeLa contaminant (HEp2), pituitary epithelial-like cell (GH3), embryonic cell (PA317), and osteosarcoma (U-2OS) cells. Of these, eight cell lines were selected: NIH-3T3, C2C12, 293T, SH-SY5Y, A549, HeLa, PA317, and U-2OS; and five underwent real-time monitoring of CDDP cytotoxicity: HeLa, A549, 293T, SH-SY5Y, and U-2OS. ATP generation was blocked 1 day after addition of 50 μM CDDP, but cytotoxicity in HeLa, A549, SH-SY5Y, and U-2OS cells could be visualized only 4 days after treatment. In 293T cells, CDDP failed to kill entirely the culture and ATP generation was only partially blocked after 1 day. This suggests potential CDDP resistance of 293T cells or metabolic clearance of the drug. Real-time monitoring and ATP measurements directly confirmed the cytotoxicity of CDDP, indicating that CDDP may interfere with mitochondrial activity.