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

IF 3.6 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Christos K Kontos, Despina Hadjichambi, Maria Papatsirou, Paraskevi Karousi, Spyridon Christodoulou, Diamantis C Sideris, Andreas Scorilas
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引用次数: 0

Abstract

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 BOK, we focused on the identification of novel BOK 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 BOK gene. Interestingly, the exons that form the back-splice junction were truncated in most circRNAs, and multiple back-splice sites were found for each BOK exon. Moreover, several BOK 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 BOK circRNAs, which could come into play in the molecular landscape and clinical investigation of ovarian and prostate cancer.

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应用纳米孔测序技术发现人卵巢和前列腺癌症细胞中凋亡相关BOK基因的新环状RNA并对其进行综合表征。
CircRNA已成为一个新的科学研究热点,越来越多的研究揭示了它们与恶性进展的关系。由于凋亡相关基因(如BOK)的circRNA存在明显的科学缺口,我们专注于在人类卵巢和前列腺癌症细胞中鉴定新的BOK circRNA。从卵巢和前列腺癌症细胞系中提取总RNA,并使用随机六聚体引物逆转录。利用基因特异性差异引物进行了一系列PCR检测。接下来,基于纳米孔技术的第三代测序,以及广泛的生物信息学分析,发现了23种新的circRNA。这些新的circRNA由BOK基因的外显子区和内含子区组成。有趣的是,在大多数circRNA中,形成背剪接连接的外显子被截短,并且每个BOK外显子都发现了多个背剪接位点。此外,一些BOK circRNA被预测为海绵状微小RNA,在生殖癌症中发挥关键作用,而假定的开放阅读框的存在表明了它们的翻译潜力。总体而言,这项研究表明,不同的选择性剪接事件导致了新的BOK circRNA的产生,这可能在卵巢和前列腺癌症的分子景观和临床研究中发挥作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Non-Coding RNA
Non-Coding RNA Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
6.70
自引率
4.70%
发文量
74
审稿时长
10 weeks
期刊介绍: Functional studies dealing with identification, structure-function relationships or biological activity of: small regulatory RNAs (miRNAs, siRNAs and piRNAs) associated with the RNA interference pathway small nuclear RNAs, small nucleolar and tRNAs derived small RNAs other types of small RNAs, such as those associated with splice junctions and transcription start sites long non-coding RNAs, including antisense RNAs, long ''intergenic'' RNAs, intronic RNAs and ''enhancer'' RNAs other classes of RNAs such as vault RNAs, scaRNAs, circular RNAs, 7SL RNAs, telomeric and centromeric RNAs regulatory functions of mRNAs and UTR-derived RNAs catalytic and allosteric (riboswitch) RNAs viral, transposon and repeat-derived RNAs bacterial regulatory RNAs, including CRISPR RNAS Analysis of RNA processing, RNA binding proteins, RNA signaling and RNA interaction pathways: DICER AGO, PIWI and PIWI-like proteins other classes of RNA binding and RNA transport proteins RNA interactions with chromatin-modifying complexes RNA interactions with DNA and other RNAs the role of RNA in the formation and function of specialized subnuclear organelles and other aspects of cell biology intercellular and intergenerational RNA signaling RNA processing structure-function relationships in RNA complexes RNA analyses, informatics, tools and technologies: transcriptomic analyses and technologies development of tools and technologies for RNA biology and therapeutics Translational studies involving long and short non-coding RNAs: identification of biomarkers development of new therapies involving microRNAs and other ncRNAs clinical studies involving microRNAs and other ncRNAs.
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