hes1-protein--human and Uveal-Neoplasms

Uveal melanoma (UM) is the most common primary intraocular tumour in adults and has a high incidence. Nearly 50% of patients with UM develop metastases after diagnosis. Long noncoding RNAs (lncRNAs) are involved in both oncogenic and tumour suppression pathways. We show that lncRNA PAUPAR is present at low levels in UM tissues and cell lines and modulates the tumourigenesis of UM in vitro and in vivo. The ectopic expression of PAUPAR in UM cells revealed that PAUPAR acts as a necessary UM suppressor and induces the silencing of HES1 expression, which significantly reduces tumour metastasis. Mechanistically, PAUPAR modulates HES1 expression by inhibiting histone H3K4 methylation. These data support a role of this lncRNA as a novel therapeutic target in cancer prevention and treatment.

Topics: Animals; Cell Line, Tumor; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Histones; Humans; Male; Melanoma; Methylation; Mice; Mice, Nude; Neoplasm Proteins; RNA, Long Noncoding; RNA, Neoplasm; Transcription Factor HES-1; Uveal Neoplasms

Controlling the spread of uveal melanoma is key to improving survival of patients with this common intraocular malignancy. The Notch ligand Jag2 has been shown to be upregulated in primary tumors that metastasize, and we therefore investigated its role in promoting invasion and clonogenic growth of uveal melanoma cells.. mRNA and protein expression of Notch pathway components were measured using qPCR and Western blot in uveal melanoma cell lines. Expression of Jag2 ligand was upregulated using Jag2-GFP-MSCV constructs or downregulated by sh-Jag2 in the uveal melanoma cell lines Mel285, Mel290, 92.1, and OMM1, and the effects on growth and invasion were assessed.. Jag2 was introduced into Mel285 and Mel290 cells, which have low baseline levels of both this ligand and Notch activity. Overall growth of the Jag2-expressing cultures increased somewhat, and a significant 3-fold increase in clonogenic growth in soft agar was also noted. Introduction of Jag2 increased motility in both wound-healing and transwell invasion assays. We also observed a significant increase in Jag2 and Hes1 mRNA in invasive OMM1 cells that had passed through a Matrigel-coated filter in the transwell assay when compared with noninvading cells. Loss-of-function studies performed in 92.1 and OMM1 lines using Jag2 shRNAs showed that downregulation of the ligand significantly suppressed cellular growth, invasion, and migration.. Our data suggest that Jag2 may play an important role in promoting Notch activity, growth, and metastasis in uveal melanoma.

Topics: Basic Helix-Loop-Helix Transcription Factors; Blotting, Western; Cell Line, Tumor; Cell Movement; Homeodomain Proteins; Humans; Intercellular Signaling Peptides and Proteins; Jagged-2 Protein; Ligands; MAP Kinase Signaling System; Melanoma; Membrane Proteins; Neoplasm Invasiveness; Neoplasm Metastasis; Phosphorylation; Real-Time Polymerase Chain Reaction; Receptors, Notch; RNA, Messenger; Signal Transduction; Transcription Factor HES-1; Up-Regulation; Uveal Neoplasms