Comparison of cancer cells in 2D vs 3D culture reveals differences in AKT-mTOR-S6K signaling and drug responses
Comparison of cancer cells in 2D vs 3D culture reveals differences in AKT-mTOR-S6K signaling and drug responses
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DOI：
10.1242/jcs.188102
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发表时间：
2017-01-01
影响因子：
4
通讯作者：
Dolznig, Helmut
中科院分区：
文献类型：
--
作者：
Riedl, Angelika;Schlederer, Michaela;Dolznig, Helmut
三维(3D)癌症模型被用作临床前系统来模拟生理性药物反应。通过对结肠癌细胞系球体的系统分析，我们从三维角度为细胞增殖和代谢能力的强烈变化提供了证据。与二维培养的细胞相比，球形细胞在AKT、哺乳动物雷帕霉素靶标(MTOR)和S6K(也称为RPS6KB1)信号通路中的活性相对较低。我们确定了信号的空间变化，因为磷酸化的RPS6水平从椭圆形表面向中心下降，这与体内血管周围的肿瘤区域密切协调。与2D模型相比，这些3D模型显示出对AKT-mTOR-S6K或丝裂原活化蛋白激酶(MAPK)通路抑制的增强的抗肿瘤反应。在2D培养中，抑制AKT-mTOR-S6K导致ERK磷酸化升高，而在此条件下，球体中ERK信号转导减少。抑制MEK1(也称为MAP2K1)导致AKT-mTOR-S6K信号在3D培养中减少，但在2D培养中不减少。这些数据表明，在3D培养和治疗期间，信号发生了明显的重新连接。血管中分离的肿瘤细胞团，除了循环中的单个肿瘤细胞外，在人类癌症的转移中起着推定的作用。因此，了解球体中的信号和3D模型中对药物治疗的反应可能有助于抗癌治疗。
Three-dimensional (3D) cancer models are used as preclinical systems to mimic physiologic drug responses. We provide evidence for strong changes of proliferation and metabolic capacity in three dimensions by systematically analyzing spheroids of colon cancer cell lines. Spheroids showed relative lower activities in the AKT, mammalian target of rapamycin (mTOR) and S6K (also known as RPS6KB1) signaling pathway compared to cells cultured in two dimensions. We identified spatial alterations in signaling, as the level of phosphorylated RPS6 decreased from the spheroid surface towards the center, which closely coordinated with the tumor areas around vessels in vivo. These 3D models displayed augmented antitumor responses to AKT-mTOR-S6K or mitogen-activated protein kinase (MAPK) pathway inhibition compared to those in 2D models. Inhibition of AKT-mTOR-S6K resulted in elevated ERK phosphorylation in 2D culture, whereas under these conditions, ERK signaling was reduced in spheroids. Inhibition of MEK1 (also known as MAP2K1) led to decreased AKT-mTOR-S6K signaling in 3D but not in 2D culture. These data indicate a distinct rewiring of signaling in 3D culture and during treatment. Detached tumor-cell clusters in vessels, in addition to circulating single tumor cells, play a putative role inmetastasis in human cancers. Hence, the understanding of signaling in spheroids and the responses in the 3D models upon drug treatment might be beneficial for anti-cancer therapies.