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鉴定人蛋白精氨酸甲基转移酶 1()基因在乳腺癌细胞中表达的新型环状 RNA

Identification of Novel Circular RNAs of the Human Protein Arginine Methyltransferase 1 () Gene, Expressed in Breast Cancer Cells.

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Biology, National and Kapodistrian University of Athens, 15701 Athens, Greece.

Laboratory of Cell Proliferation & Ageing, Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos", 15341 Aghia Paraskevi, Greece.

出版信息

Genes (Basel). 2022 Jun 24;13(7):1133. doi: 10.3390/genes13071133.

Abstract

Circular RNAs (circRNAs) constitute a type of RNA formed through back-splicing. In breast cancer, circRNAs are implicated in tumor onset and progression. Although histone methylation by PRMT1 is largely involved in breast cancer development and metastasis, the effect of circular transcripts deriving from this gene has not been examined. In this study, total RNA was extracted from four breast cancer cell lines and reversely transcribed using random hexamer primers. Next, first- and second-round PCRs were performed using gene-specific divergent primers. Sanger sequencing followed for the determination of the sequence of each novel circRNA. Lastly, bioinformatics analysis was conducted to predict the functions of the novel circRNAs. In total, nine novel circRNAs were identified, comprising both complete and truncated exons of the gene. Interestingly, we demonstrated that the back-splice junctions consist of novel splice sites of the exons. Moreover, the circRNA expression pattern differed among these four breast cancer cell lines. All the novel circRNAs are predicted to act as miRNA and/or protein sponges, while five circRNAs also possess an open reading frame. In summary, we described the complete sequence of nine novel circRNAs of the gene, comprising distinct back-splice junctions and probably having different molecular properties.

摘要

环状 RNA(circRNAs)是通过反向剪接形成的一类 RNA。在乳腺癌中,circRNAs 参与肿瘤的发生和发展。尽管 PRMT1 介导的组蛋白甲基化在很大程度上参与了乳腺癌的发展和转移,但该基因衍生的环状转录本的影响尚未被检测到。在这项研究中,从四种乳腺癌细胞系中提取总 RNA,并使用随机六聚体引物进行逆转录。接下来,使用基因特异性发散引物进行第一和第二轮 PCR。随后进行 Sanger 测序以确定每个新 circRNA 的序列。最后,进行生物信息学分析以预测新 circRNAs 的功能。总共鉴定出了 9 种新型 circRNAs,它们包含基因的完整和截断外显子。有趣的是,我们证明了反向剪接接头由基因外显子的新剪接位点组成。此外,circRNA 的表达模式在这四种乳腺癌细胞系中存在差异。所有新的 circRNAs 被预测为 miRNA 和/或蛋白质海绵,而五个 circRNAs 也具有开放阅读框。总之,我们描述了基因的 9 种新型 circRNAs 的完整序列,它们包含不同的反向剪接接头,并且可能具有不同的分子特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6df/9316507/45bce822dab3/genes-13-01133-g001.jpg

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