Silencing of ANKRD12 circRNA induces molecular and functional changes associated with invasive phenotypes
| Autor: | Wafa Al Ameri, Arash Rafii, Joel A. Malek, Yasmin Ali Mohamoud, Simeon S. Andrews, Fatima M. Al-Dasim, Thasni Karedath, Samson Mathews Samuel, Ikhlak Ahmed, Iman K. Al-Azwani |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cancer Research Lung Neoplasms RNA-Seq Exon 0302 clinical medicine Breast cancer Cell Movement Cyclin D1 Breast RNA Small Interfering Lung Nuclear Proteins Exons Cell cycle lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens Phenotype OXPHOS Cell biology Gene Expression Regulation Neoplastic Oncology 030220 oncology & carcinogenesis MCF-7 Cells Research Article RNase R Cancer invasion Breast Neoplasms Biology Transfection lcsh:RC254-282 03 medical and health sciences Circular RNA Genetics medicine Biomarkers Tumor Gene silencing Humans Neoplasm Invasiveness Gene Silencing Gene Messenger RNA Cancer RNA Circular medicine.disease G1 Phase Cell Cycle Checkpoints OCR 030104 developmental biology siRNA Cancer cell RNA-seq |
| Zdroj: | BMC Cancer, Vol 19, Iss 1, Pp 1-17 (2019) BMC Cancer |
| DOI: | 10.1101/366245 |
| Popis: | Background Circular RNAs (circRNAs) that form through non-canonical backsplicing events of pre-mRNA transcripts are evolutionarily conserved and abundantly expressed across species. However, the functional relevance of circRNAs remains a topic of debate. Methods We identified one of the highly expressed circRNA (circANKRD12) in cancer cell lines and characterized it validated it by Sanger sequencing, Real-Time PCR. siRNA mediated silencing of the circular junction of circANKRD12 was followed by RNA Seq analysis of circANKRD12 silenced cells and control cells to identify the differentially regulated genes. A series of cell biology and molecular biology techniques (MTS assay, Migration analysis, 3D organotypic models, Real-Time PCR, Cell cycle analysis, Western blot analysis, and Seahorse Oxygen Consumption Rate analysis) were performed to elucidate the function, and underlying mechanisms involved in circANKRD12 silenced breast and ovarian cancer cells. Results In this study, we identified and characterized a circular RNA derived from Exon 2 and Exon 8 of the ANKRD12 gene, termed here as circANKRD12. We show that this circRNA is abundantly expressed in breast and ovarian cancers. The circANKRD12 is RNase R resistant and predominantly localized in the cytoplasm in contrast to its source mRNA. We confirmed the expression of this circRNA across a variety of cancer cell lines and provided evidence for its functional relevance through downstream regulation of several tumor invasion genes. Silencing of circANKRD12 induces a strong phenotypic change by significantly regulating cell cycle, increasing invasion and migration and altering the metabolism in cancer cells. These results reveal the functional significance of circANKRD12 and provide evidence of a regulatory role for this circRNA in cancer progression. Conclusions Our study demonstrates the functional relevance of circANKRD12 in various cancer cell types and, based on its expression pattern, has the potential to become a new clinical biomarker. Electronic supplementary material The online version of this article (10.1186/s12885-019-5723-0) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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