Comparison of 2D-and 3D-culture models as drug-testing platforms in breast cancer
Comparison of 2D-and 3D-culture models as drug-testing platforms in breast cancer
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DOI：
10.3892/or.2015.3767
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发表时间：
2015-04-01
期刊：
影响因子：
4.2
通讯作者：
Minami, Hironobu
中科院分区：
文献类型：
--
作者：
Imamura, Yoshinori;Mukohara, Toru;Minami, Hironobu
人们逐渐认识到，在使用二维（2D）培养细胞系的平台上筛选肿瘤药物无法精确选择临床活性药物；因此，三维（3D）培养系统正在兴起，并显示出更好地模拟体内肿瘤微环境的潜力。本研究的目的是揭示化疗药物在 2D 和 3D 培养物之间的差异效应，并探讨其潜在机制。我们通过评估药物敏感性、氧状态以及 Ki-67 和半胱天冬酶的表达来评估 2D 和 3D 培养的乳腺癌细胞系之间的差异。三种细胞系（BT-549、BT-474 和 T-47D）在 3D 培养中形成致密的多细胞球体 (MCS)，并且与 2D 培养的细胞相比，对紫杉醇和阿霉素表现出更大的耐药性。另外三种细胞系（MCF-7、HCC-1954 和 MDA-MB-231）在 3D 培养中仅形成松散的 MCS，并且表现出与 2D 培养中发现的药物敏感性相似的药物敏感性。与 3D 培养物相比，紫杉醇处理导致 2D 培养物中 cleaved-PARP 表达的增加更大，但仅在形成致密 3D-MCS 的细胞系中，表明 MCS 的形成保护细胞免受紫杉醇诱导的细胞凋亡。仅在致密的 3D-MCS 中观察到缺氧。与 2D 培养相比，BT-549 在 3D 培养中的 Ki-67 阳性细胞较少，这表明更多的 GO 休眠亚群是其在 3D 培养中产生耐药性的原因。 BT-474 在 3D 培养中的 caspase-3 水平低于 2D 培养中的 caspase-3 水平，表明 3D 环境具有抗凋亡作用。最后，我们比较了源自患者异种移植物 (PDX)、新鲜 PDX 肿瘤和患者原始肿瘤的 2D 和 3D 原代培养细胞中 Ki-67 和 caspase 的染色情况； 2D 培养的细胞显示出更大比例的 Ki-67 阳性和 caspase-3 阳性细胞，这与 3D 原代培养更好地代表体内肿瘤特征的观点一致。总之，形成致密 MCS 的 3D 培养细胞可能比 2D 培养细胞更好地模拟体内重要的肿瘤特征，即缺氧、休眠、抗凋亡特征及其产生的耐药性。
It is becoming recognized that screening of oncology drugs on a platform using two-dimensionally (2D)-cultured cell lines is unable to precisely select clinically active drugs; therefore three-dimensional (3D)-culture systems are emerging and show potential for better simulating the in vivo tumor microenvironment. The purpose of this study was to reveal the differential effects of chemotherapeutic drugs between 2D- and 3D-cultures and to explore their underlying mechanisms. We evaluated differences between 2D- and 3D-cultured breast cancer cell lines by assessing drug sensitivity, oxygen status and expression of Ki-67 and caspases. Three cell lines (BT-549, BT-474 and T-47D) developed dense multicellular spheroids (MCSs) in 3D-culture, and showed greater resistance to paclitaxel and doxorubicin compared to the 2D-cultured cells. An additional three cell lines (MCF-7, HCC-1954, and MDA-MB-231) developed only loose MCSs in 3D, and showed drug sensitivities similar to those found in the 2D-culture. Treatment with paclitaxel resulted in greater increases in cleaved-PARP expression in the 2D-culture compared with the 3D-culture, but only in cell lines forming dense 3D-MCSs, suggesting that MCS formation protected the cells from paclitaxel-induced apoptosis. Hypoxia was observed only in the dense 3D-MCSs. BT-549 had fewer cells positive for Ki-67 in 3D-than in 2D-culture, suggesting that the greater GO-dormant subpopulation was responsible for its drug resistance in the 3D-culture. BT-474 had a lower level of caspase-3 in the 3D-than in the 2D-culture, suggesting that the 3D-environment was anti-apoptotic. Finally, we compared staining for Ki-67 and caspases in the 2D- and 3D-primary-cultured cells originating from a patient-derived xenograft (PDX), fresh PDX tumor, and the patient's original tumor; 2D-cultured cells showed greater proportions of Ki-67-positive and caspase-3-positive cells, in agreement with the view that 3D-primary culture better represents characteristics of tumors in vivo. In conclusion, 3D-cultured cells forming dense MCSs may be better than 2D-cultured cells in simulating important tumor characteristics in vivo, namely hypoxia, dormancy, anti-apoptotic features and their resulting drug resistance.