Project description:MEG3 was known as a growth suppressor in tumor cells by activating p53. Besides, MEG3 could regulate transforming growth factor-β (TGF-β) signaling pathway, which is the key regulator of skeletal myogenesis and can enhance the proliferation of myogenic cells. Previous study also showed MEG3 was highly expressed in the paraxial mesoderm and probably regulated muscle development. To investigate the potential function of MEG3 in muscle development, we detected the expression levels of protein-coding genes after MEG3 over-expression or knockdown in C2C12 cell line using microarrays.
Project description:To investigate the collective role of the Dlk1-Dio3 ncRNA cluster in myogenesis, we developed two stable CRISPR-Cas9 gene-edited C2C12 skeletal muscle cell lines, Meg3-proximal cis-acting sequence (Meg3-CAS or 2-4) to over-express and Meg3-transcription start site (Meg3-TSS or 2-7) to under-express the entire mega-cluster of Dlk1-Dio3 ncRNAs. Mutations affecting the dosage of all ncRNAs expressed from the Dlk1-Dio3 locus cause profound changes in myoblast fusion and differentiation. To evaluate genome-wide perturbations in chromatin accessibility we evaluated specific histone lysine modifications, inducing the repressive histone mark, H3K27me3, and the activating mark, H3K4me3
Project description:To investigate the collective role of the Dlk1-Dio3 ncRNA cluster in myogenesis, we developed two stable CRISPR-Cas9 gene-edited C2C12 skeletal muscle cell lines, Meg3-proximal cis-acting sequence (Meg3-CAS or 2-4) to over-express and Meg3-transcription start site (Meg3-TSS or 2-7) to under-express the entire mega-cluster of Dlk1-Dio3 ncRNAs. Mutations affecting the dosage of all ncRNAs expressed from the Dlk1-Dio3 locus cause profound changes in myoblast fusion and differentiation. To evaluate genome-wide perturbations in chromatin accessibility we evaluated specific histone lysine modifications, inducing the repressive histone mark, H3K27me3, and the activating mark, H3K4me3
Project description:Hepatoblastoma is the most common primary liver malignancy in children, and outcomes remain poor for patients with metastatic disease. Long non-coding RNAs (lncRNAs) regulate tumor behavior, but their role in metastatic hepatoblastoma is not well defined. This study investigated the expression and functional significance of the lncRNA, maternally expressed gene 3 (MEG3), in a metastatic model of hepatoblastoma. RNA sequencing compared the metastatic hepatoblastoma cell line HLM_2 with its parental HuH6 cell line to identify differentially expressed lncRNAs. MEG3 expression was examined using hepatoblastoma patient datasets and validated using qPCR in cell lines, orthotopic tumors, and COA67 patient-derived xenografts. Functional effects of MEG3 knockdown in HLM_2 cells were evaluated with siRNA to evaluate clonogenicity, tumorsphere formation, migration, and invasion. The effects of MEG3 overexpression on clonogenicity, migration, and invasion were assessed in HuH6 cells. MEG3 was significantly upregulated in metastatic cells and orthotopic tumors compared with controls. MEG3 silencing reduced clonogenicity, tumorsphere formation, migration, and invasion. MEG3 overexpression increased clonogenicity, migration, and invasion. These findings indicate that MEG3 is upregulated in metastatic hepatoblastoma and may contribute to aggressive tumor phenotype, warranting further examination of the mechanistic role of MEG3 in hepatoblastoma, and its relevance as a biomarker or therapeutic target.
Project description:This experiment was designed to detect differences in PRC2 occupancy in a human induced pluripotent cell line (MRC5) naturally deficient in MEG3 RNA after lentiviral overexpression of MEG3. MRC5 were transduced with lentiviruses for the overexpression of MEG3 lncRNA or GFP as a control. ChIP-seq was performed with EZH2 and JARID2 antibodies.
