Endothelial Pim3 kinase protects the vascular barrier during lung metastasis

To investigate transcriptome changes of endothelial cells during cancer cell response, we studied HUVEC RNA expression in conditioned medium derived from empty plates versus B16-F10 melanoma cultures. Endothelial cells (ECs) form a tissue-specific barrier for disseminating cancer cells in distant organs. However, the molecular regulation of the ECs in the metastatic niche is poorly understood. Here, we analyzed the transcriptional reprogramming of cultured human ECs (HUVECs) in response to potential signal mediators sectered to the cultured medium by B16-F10 melanoma cells. Melanoma cells were cultured for 48h, after which conditioned medium was collected, spun and added to replace HUVEC culture medium. As a control, HUVEC cultures were treated with conditioned medium stored in similar conditions on empty plates. HUVEC cells were cultured in conditioned medium for 16 h after which total RNA was isolated and processed for mRNA-Seq.