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CircEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration [qRT-PCR validation]

ABSTRACT: circEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration [qRT-PCR validation]

PROVIDER: PRJNA781249 | ENA |

REPOSITORIES: ENA

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circEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration [qRT-PCR validation]

Project description:Background: Hepatocellular carcinoma (HCC) is one kind of the leading causes of cancer-related deaths in the world. Recent evidence indicates that circular RNAs (circRNAs) play important roles in tissue development, gene transcription, signal regulation and tumorigenesis. However, whether circRNAs are involved in HCC progression and encode functional proteins remains largely unknown. Methods: circRNAs microarrays were performed using pathologically diagnosed HCC samples and paired adjacent normal liver tissues. Cell invasion, migration, cycle, and apoptosis post circRNA overexpression were measured using a transwell culture system, a wound healing assay, and flow cytometry. Full-length, mutated, and truncated sequences of circEPS15 with a FLAG tag were inserted into a circular expression vector. Western blotting was used to confirm circEPS15 expression and the requirement of internal ribosomal entry site (IRES) elements within the circRNA. The miRNA and mRNA expression profiles were obtained by analyzing data retrieved from The Cancer Genome Atlas (TCGA) database. We then constructed a ceRNA network ofcircEPS15, miRNAs, and mRNAs. Results: The expression of circEPS15 was downregulated in HCC tissues, and the survival curves showed that low circEPS15 levels were associated with poor overall survival in HCC patients. Overexpression of circEPS15 suppressed tumor invasion and migration by inhibiting the TJP1/CDH2/VIM signaling pathway and retarded cell cycle progression, yet had no effect on cell apoptosis. ceRNA analysis and qRT-PCR results suggest a possible circRNA (circEPS15)-miRNA-mRNA network in HCC. The spanning junction open reading frame in circEPS15 driven by IRES encoded a novel protein. Conclusions: Endogenous circEPS15 plays a novel role in repressing HCC through the ceRNA network and encoding a functional protein.

2021-11-19 | GSE189043 | GEO

circEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration

Project description:This study evaluates the effect circular RNA (circRNA) has on the development of hepatocellular carcinoma (HCC), which has been an area of limited information in the field. Through microarray analysis, we identified and then characterized the circRNA, circEPS15, which is downregulated in HCC tissue compared to noncancerous tissues. Our analysis shows that overexpressing circEPS15 could reduce tumor cell migration through the inhibition of cell cycle signaling pathways, suggesting this circRNA could be a target for novel HCC therapies. Moreover, we show that circEPS15 encodes a functional protein that could contribute to its antitumor effects. We believe that our study makes a significant contribution to the literature because it both clarifies the molecular mechanisms of circRNA in cell growth and development, as well as provides novel marker or target candidates for improving HCC diagnosis and treatment.

2021-01-15 | GSE164803 | GEO

circEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration

Project description:circEPS15 overexpression in hepatocellular carcinoma modulates tumor invasion and migration

| PRJNA692101 | ENA

circEPS15 Overexpression in Hepatocellular Carcinoma Modulates Tumor Invasion and Migration.

Project description:Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Recent evidence has shown that circular RNAs (circRNAs) play important roles in tissue development, gene transcription, signal regulation and tumorigenesis. However, whether circRNAs are involved in HCC progression and encode functional proteins remains largely unknown. In the present study, we aimed to explore the function and molecular mechanism of circRNAs in HCC. First, many circRNAs were found to be differentially expressed in HCC samples and paired adjacent normal liver tissues. The validation of dysregulated circRNAs by qRT-PCR revealed that circEPS15 expression was downregulated in HCC tissues, and the survival curves showed that low circEPS15 levels were associated with poor overall survival in HCC patients. Then, the overexpression of circEPS15 suppressed tumor cell invasion and migration by inhibiting the TJP1/CDH2/VIM signaling pathway and retarded cell cycle progression, which was confirmed by the Transwell culture system, wound healing assays, flow cytometry and western blot assays. After that, the spanning junction open reading frame in circEPS15 driven by IRES was shown to encode a novel protein, which was verified by western blotting with full-length, mutated, and truncated sequences of circEPS15 with a FLAG tag. Moreover, ceRNA analysis and qRT-PCR results suggest a possible circRNA (circEPS15)-miRNA-mRNA network in HCC. Collectively, our study reveals that endogenous circEPS15 plays a novel role in repressing HCC through the ceRNA network and encodes a functional protein.

| S-EPMC8861492 | biostudies-literature

miRNA overexpression modulates neutrophil migration

Project description:miRNA overexpression modulates neutrophil migration

| PRJNA704949 | ENA

Agreement in Breast Cancer Classification between Microarray and qRT-PCR from Fresh-Frozen and Formalin-Fixed Tissues

Project description:Determining agreement in classification between platforms and procurement methods requires a variety of methods. We have shown that centroid-based algorithms are robust classifiers for breast cancer subtype assignment across platforms (microarray and qRT-PCR data) and procurement conditions (fresh frozen and formalin-fixed, paraffin-embedded tissues). On a gene-by-gene basis, we found that the standard deviation, dynamic range, and concordance correlation coefficient are important parameters to assess individual primer set performance across procurement methods. Our strategy for primer set validation and classification have applications in routine clinical practice for stratifying breast cancers and other tumor types Keywords: reference x sample

2007-04-20 | GSE6130 | GEO

Genome wide (GW) and validation CRISPR screens on encephalitogenic CD4+ T cell migration into the CNS

Project description:To identify genes driving encephalitogenic CD4+ T cell migration into the CNS, we performed a genome-wide CRISPR screen and a subsequent validation screen For the genome-wide CRISPR screen, up to 4 sgRNA per gene and 800 non targeted controls were included, for a total of 87690 individual sgRNAs, and for the validation screen, up to 6 sgRNA per gene and 241 non targeted controls for a total of 12000 individual sgRNAs

2023-08-01 | GSE232340 | GEO

miR-99 overexpression modulates neutrophil migration

Project description:miR-99 overexpression modulates neutrophil migration

| PRJNA605187 | ENA

Technical comparison of qRT-PCR experiments with the GLYCOv2 custom array

Project description:Gene expression patterns were surveyed using RNA from 4 different murine sources. Four classes of RNA were analyzed: undifferentiated embryonic stem cells, differentiated ES cells, kidney and liver tissues. Classes were prepared in triplicate for a total of 12 samples. All samples were hybridized to the custom designed CFG GLYCOv2 glycogene array. Analysis results were used for comparison with parallel experiments using qRT-PCR.

2011-03-02 | E-GEOD-27595 | biostudies-arrayexpress

Real-time quantitative PCR analysis for validation of miRNAs expressed during osteogenic differentiation of bone marrow MSCs versus healthy donors MSCs

Project description:Total small RNA was isolated from BM-MSCs obtained from OA diagnosed patients (n=10) and non-OA (n=10 after osteogenic induction for 1, 10 and 21 days. miRNAs, were initially screened and those with consistent differential expression were selected for further validation by qRT-PCR analysis. From an initial screening of 246 differentially expressed miRNAs (fold change ?± 2, P ?0.05) between OA and non-OA BM-MSC samples 21 were selected and validated by qRT-PCR

2015-07-14 | E-GEOD-70856 | biostudies-arrayexpress

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