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Open Access 01-12-2019 | Breast Cancer | Research article

Silencing of ANKRD12 circRNA induces molecular and functional changes associated with invasive phenotypes

Authors: Thasni Karedath, Ikhlak Ahmed, Wafa Al Ameri, Fatima M. Al-Dasim, Simeon S. Andrews, Samson Samuel, Iman K. Al-Azwani, Yasmin Ali Mohamoud, Arash Rafii, Joel A. Malek

Published in: BMC Cancer | Issue 1/2019

Abstract

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.

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Title: Silencing of ANKRD12 circRNA induces molecular and functional changes associated with invasive phenotypes
Authors: Thasni Karedath
Ikhlak Ahmed
Wafa Al Ameri
Fatima M. Al-Dasim
Simeon S. Andrews
Samson Samuel
Iman K. Al-Azwani
Yasmin Ali Mohamoud
Arash Rafii
Joel A. Malek
Publication date: 01-12-2019
Publisher: BioMed Central
Keywords: Breast Cancer
Breast Cancer
Published in: BMC Cancer / Issue 1/2019
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-019-5723-0

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Abstract

Background

Methods

Results

Conclusions

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