Discovery and Comprehensive Characterization of Novel Circular RNAs of the Apoptosis-Related <i>BOK</i> Gene in Human Ovarian and Prostate Cancer Cells, Using Nanopore Sequencing

CircRNAs have become a novel scientific research hotspot, and an increasing number of studies have shed light on their involvement in malignant progression. Prompted by the apparent scientific gap in circRNAs from apoptosis-related genes, such as <i>BOK</i>, we focused on the identificat...

Full description

Bibliographic Details
Main Authors: Christos K. Kontos, Despina Hadjichambi, Maria Papatsirou, Paraskevi Karousi, Spyridon Christodoulou, Diamantis C. Sideris, Andreas Scorilas
Format: Article
Language:English
Published: MDPI AG 2023-09-01
Series:Non-Coding RNA
Subjects:
Online Access:https://www.mdpi.com/2311-553X/9/5/57
Description
Summary:CircRNAs have become a novel scientific research hotspot, and an increasing number of studies have shed light on their involvement in malignant progression. Prompted by the apparent scientific gap in circRNAs from apoptosis-related genes, such as <i>BOK</i>, we focused on the identification of novel <i>BOK</i> circRNAs in human ovarian and prostate cancer cells. Total RNA was extracted from ovarian and prostate cancer cell lines and reversely transcribed using random hexamer primers. A series of PCR assays utilizing gene-specific divergent primers were carried out. Next, third-generation sequencing based on nanopore technology followed by extensive bioinformatics analysis led to the discovery of 23 novel circRNAs. These novel circRNAs consist of both exonic and intronic regions of the <i>BOK</i> gene. Interestingly, the exons that form the back-splice junction were truncated in most circRNAs, and multiple back-splice sites were found for each <i>BOK</i> exon. Moreover, several <i>BOK</i> circRNAs are predicted to sponge microRNAs with a key role in reproductive cancers, while the presence of putative open reading frames indicates their translational potential. Overall, this study suggests that distinct alternative splicing events lead to the production of novel <i>BOK</i> circRNAs, which could come into play in the molecular landscape and clinical investigation of ovarian and prostate cancer.
ISSN:2311-553X