Project description:Prognosis of non small cell lung cancer (NSCLC) is very poor mainly because it often has metastasized to distant organs already at the time of diagnosis. Therefore biomarkers which can predict metastasis are urgently needed. miRNAs have been shown to play important roles in the regulation of different tumor cell processes including those involved in metastasis. We recently showed that miRNA-214 is linked to a radioresistant phenotype of NSCLC. Interestingly, altered miRNA-214 expression has in breast, cervical and melanoma been linked to metastasis. We therefore examined the role of miRNA-214 in the metastasis of NSCLC cells. We found that down regulation of miRNA-214 increased invasive potential of NSCLC cells and conversely, overexpression of miRNA-214 in NSCLC cells with low endogenous level of miRNA-214, decreased invasiveness. Gene expression analyses of NSCLC cells with low and high levels of miRNA-214, followed by bioinformatics analyses identified a number of target genes which were linked to metastasis including pregnancy associated plasma protein A (PAPP-A), alpha protein kinase 2 (ALPK2), cyclin-dependent kinase 6 (CDK6) and tumor necrosis-factor alpha-induced protein 3 (TNFAIP3). These targets were validated on mRNA and protein level. U-1810 cells with high endogenous level of miRNA-214 were treated with miRNA-214 antagomir (3 biological replicates) or non-targeting antagomir (3 biological replicates) and then RNA was extracted and applied to Affymetrix gene array platform.
Project description:Prognosis of non small cell lung cancer (NSCLC) is very poor mainly because it often has metastasized to distant organs already at the time of diagnosis. Therefore biomarkers which can predict metastasis are urgently needed. miRNAs have been shown to play important roles in the regulation of different tumor cell processes including those involved in metastasis. We recently showed that miRNA-214 is linked to a radioresistant phenotype of NSCLC. Interestingly, altered miRNA-214 expression has in breast, cervical and melanoma been linked to metastasis. We therefore examined the role of miRNA-214 in the metastasis of NSCLC cells. We found that down regulation of miRNA-214 increased invasive potential of NSCLC cells and conversely, overexpression of miRNA-214 in NSCLC cells with low endogenous level of miRNA-214, decreased invasiveness. Gene expression analyses of NSCLC cells with low and high levels of miRNA-214, followed by bioinformatics analyses identified a number of target genes which were linked to metastasis including pregnancy associated plasma protein A (PAPP-A), alpha protein kinase 2 (ALPK2), cyclin-dependent kinase 6 (CDK6) and tumor necrosis-factor alpha-induced protein 3 (TNFAIP3). These targets were validated on mRNA and protein level.
Project description:MicroRNAs have been demonstrated to be deregulated in multiple myeloma (MM). We have previously reported the downregulation of miR-214 in MM compared to normal plasma cells. In the present study, we have explored the functional role of miR-214 in myeloma pathogenesis. Ectopic expression of miR-214 reduced cell growth and induced apoptosis of myeloma cells. In order to identify the potential direct target genes of miR-214 which could be involved in the biological pathways regulated by this miRNA, gene expression profiling of H929 myeloma cell line transfected with precursor miR-214 was carried out. Functional analysis revealed significant enrichment for DNA replication, cell cycle phase and DNA binding. We show that miR-214 directly down-regulates the expression of PSMD10, which encodes the oncoprotein gankyrin, and ASF1B, a histone chaperone required for DNA replication, by binding to their 3'-UTR. In addition, gankyrin inhibition induced an increase of P53 mRNA levels and subsequent up-regulation in CDKN1A (p21Waf1/Cip1) and BAX transcripts, which are direct transcriptional targets of p53. In conclusion, we demonstrate that miR-214 function as a tumor suppressor in myeloma by a positive regulation of p53 and inhibition of DNA replication. H929 cell line was transfected with Pre-miR™ miRNA precursors pre-miR-214 or pre-miR™ miRNA negative, non-targeting control#1 (Ambion) at 50 nM concentration, using the nucleofector II system with C-16 program (Amaxa). The experiments were performed in triplicates.
Project description:The small RNA landscape of pediatric ALK-positive ALCL was defined by RNA sequencing. Overall, 121 miRNAs were significantly dysregulated in ALCL compared to non-neoplastic lymph nodes. The most up-regulated miRNA was miR-21-5p, while miR-19a-3p and miR-214-5p were reduced in ALCL. Characterization of miRNA expression in cases that relapsed after first line therapy disclosed a significant association between miR-214-5p down-regulation and aggressive non-common histology.
Project description:Illumina miRNA-seq method to uncover the expression profile of NSCLC in-vitro experimental models consisting of cell lines A549, H460 compared to healthy BEAS-2B cell line, and lung tissue (NSCLC and paired normal) from urethane treated 6-week-old FVB/NJ mice. We aimed to uncover the divergent epigenetic background of KRAS-mutant NSCLC in mouse and human cell lines, extensively used as biological models in relevant research. To that end, we have comprehensively mapped the functional miRNA and lncRNA landscape of human (A549 and H460) and mouse (experimentally developed LUAD) NSCLC models and correlated current results with LRF/ZBTB7A expression
Project description:Ongoing immunomodulatory strategies in tumors characterized by an overall hot immune phenotype may improve prognosis of patients with non-small cell lung cancer (NSCLC). Our objective was to develop a reliable and stable scoring system for the identification of immunologically hot NSCLC and to evaluate its association with response to immunotherapies. A Hot Oral Tumor (HOT) score was developed using data from The Cancer Genome Atlas. HOT score was computed in 82 patients with NSCLC treated with second-line immunotherapy targeting PD-1/PD-L1. High HOT score was associated with a statistically significant improved clinical outcome.
Project description:Abstract: Background & Aims: Unusual hypervascularity is a hallmark of human hepatocellular carcinoma (HCC). Although microRNA-214 (miR-214) is upregulated in other human cancers, it is downregulated in HCC. We elucidated the biological and clinical significance of miR-214 downregulation in HCC. Methods: MicroRNAs deregulated in HCC were identified using array-based MicroRNA profiling. A luciferase reporter assay confirmed target association between miR-214 and hepatoma-derived growth factor (HDGF). Tube formation and in vivo angiogenesis assays validated the roles of miR-214/HDGF in angiogenesis. Results: MiR-214 downregulation was associated with higher tumor recurrence and worse clinical outcomes. Ectopic expression of miR-214 suppressed xenograft tumor growth and microvascularity of the tumor and its surrounding tissues. The genes downregulated by ectopic expression of miR-214 were involved in the regulation of apoptosis, cell cycle, and angiogenesis. Integrated analysis disclosed HDGF as a downstream target of miR-214. Conditioned medium of HCC cells contained bioactivity to stimulate tube formation of human umbilical vein endothelial cells, which was abolished by pretreatment of the conditioned media with HDGF antibodies, silencing of HDGF expression or ectopic expression of miR-214 in the donor HCC cells. The angiogenic activity of the conditioned media lost by ectopic expression of miR-214 in the donor cells was restored by supplementation with recombinant HDGF. In vivo tumor angiogenesis assays showed significant suppression of tumor vascularity by ectopic expression of miR-214. Conclusions: A novel role of microRNA in tumrigenesis is identified. Downregulation of miR-214 contributes to unusual hypervascularity of HCC via activation of the HDGF paracrine pathway for tumor angiogenesis. To identify miRNAs that are deregulated in human HCC, 68 HCC and 21 non-tumor liver tissues were subjected to profiling of miRNA expression using miRNA arrays containing 739 human miRNA probes. Differentially expressed microRNAs were identified.
Project description:MicroRNAs (miRNAs) are small non-coding RNAs that act as post-transcriptional gene modulators. Ginsenoside-Rg1, one of the active components of ginseng, has been confirmed by us as an angiogenesis inducer. Using miRNA microarray analysis, a total of 15 (including miR-214) and 3 miRNAs were found to be down- or up-regulated by Rg1 in human umbilical vein endothelial cells (HUVEC), respectively. Since miR-214 is closely related to endothelial nitric oxide synthase (eNOS) and hence angiogenesis; its expression was further validated by qRT-PCR. We also investigated the role of miR-214 on eNOS expression and in tubulogenesis of HUVEC by transfection of specific miRNA inhibitor or precursor. Our results suggested that Rg1 can down-regulate miR-214 expression in HUVEC, leading to an increase in eNOS expression which can promote angiogenesis. This result signifies a new understanding towards how a simple natural compound can affect physiological changes through modulation of miRNA expression. The study is used to investigate the role of miRNA-214 in Rg1-induced human endothelial cells.
Project description:To identify radiation-induced miRNAs, we initially profiled human miRNA expression in NSCLC A549 and H1299 cells treated with X-ray radiation using miRNA microarrays. Indeed, we observed multiple dysregulated miRNAs following radiation in NSCLC cell lines.
Project description:We developed a strategy to generate cardiac progenitor cells from human induced pluripotent stem cells using a novel small molecule.miRNA-sequencing results showed different miRNA expression profile among undifferentiated human induced pluripotent stem cells(hiPSCs), DMSO and ISX-9 treated hiPSCs.In comparsion with DMSO treated cells, ISX-9 upregulated several myogenic miRNAs and cardiac hypertrophy related-miRNAs including miR-335, miR-21, miR-30c,miRNA-181a and miR-214.