RNAi screens for Rho GTPase regulators of cell shape and YAP/TAZ localisation in triple negative breast cancer.
Patricia Pascual-VargasSamuel CooperJulia SeroVicky BousgouniMar Arias-GarciaChris BakalPublished in: Scientific data (2017)
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.
Keyphrases
- single cell
- induced apoptosis
- protein kinase
- cell cycle arrest
- cell therapy
- squamous cell carcinoma
- small cell lung cancer
- genome wide
- high throughput
- magnetic resonance imaging
- transcription factor
- cell death
- oxidative stress
- copy number
- computed tomography
- stem cells
- endoplasmic reticulum stress
- pi k akt
- genome wide analysis
- bioinformatics analysis