Dynamical Cell Systems Team, Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
Department of Computational Systems Medicine, Imperial College London, South Kensington Campus, London SW7, UK.
Sci Data. 2017 Mar 1;4:170018. doi: 10.1038/sdata.2017.18.
In order to metastasise, triple negative breast cancer (TNBC) must make dynamic changes in cell shape. The shape of all eukaryotic cells is regulated by Rho Guanine Nucleotide Exchange Factors (RhoGEFs), which activate Rho-family GTPases in response to mechanical and informational cues. In contrast, Rho GTPase-activating proteins (RhoGAPs) inhibit Rho GTPases. However, which RhoGEFs and RhoGAPS couple TNBC cell shape to changes in their environment is very poorly understood. Moreover, whether the activity of particular RhoGEFs and RhoGAPs become dysregulated as cells evolve the ability to metastasise is not clear. Towards the ultimate goal of identifying RhoGEFs and RhoGAPs that are essential for TNBC metastasis, we performed an RNAi screen to isolate RhoGEFs and RhoGAPs that contribute to the morphogenesis of the highly metastatic TNBC cell line LM2, and its less-metastatic parental cell line MDA-MB-231. For ~6 million cells from each cell line, we measured 127 different features following the depletion of 142 genes. Using a linear classifier scheme we also describe the morphological heterogeneity of each gene-depleted population.
为了转移，三阴性乳腺癌（TNBC）必须在细胞形状上发生动态变化。所有真核细胞的形状都受 Rho 鸟嘌呤核苷酸交换因子（RhoGEFs）的调节，这些因子根据机械和信息线索激活 Rho 家族 GTPases。相比之下，Rho GTPase 激活蛋白（RhoGAPs）抑制 Rho GTPases。然而，哪些 RhoGEFs 和 RhoGAPs 将 TNBC 细胞形状与环境变化联系起来，目前还知之甚少。此外，当细胞进化出转移能力时，特定 RhoGEFs 和 RhoGAPs 的活性是否失调尚不清楚。为了最终确定对 TNBC 转移至关重要的 RhoGEFs 和 RhoGAPs，我们进行了 RNAi 筛选，以分离有助于高度转移性 TNBC 细胞系 LM2 及其转移性较低的亲本细胞系 MDA-MB-231 形态发生的 RhoGEFs 和 RhoGAPs。对于来自每个细胞系的约 600 万个细胞，我们在耗尽 142 个基因后测量了 127 个不同的特征。我们还使用线性分类器方案描述了每个基因耗尽群体的形态异质性。
