Identification of Circular RNAs Altered in Mouse Jejuna After Radiation

Abstract

Background/Aims: Circular RNAs (circRNAs) make up a large class of non-coding RNAs and play important roles in a variety of diseases, including nervous system diseases and cancers. The intestinal epithelium is sensitive to ionizing radiation, radiotherapy of abdominal or pelvic tumors or nuclear accident exposure can lead to high radiation toxicity, which can result in radiation-induced intestinal injury. The goal of this present study was to analyze the potential roles of circRNAs in radiation-induced intestinal injury. Methods: Mice were divided into two groups: control group and irradiated group. Irradiated group was 3.5 days after 14Gy abdominal irradiation (ABI) group. We started with RNA-seq of circRNA changes in mouse jejuna after radiation and validated by RT-PCR in the following experimental. miRNAs targeted mRNAs were predicted using proprietary software based on target scan and Miranda. The network of circRNA-miRNA-mRNA was illustrated by cytoscape software. Results: 2751 circRNAs were detected in the two groups. At day 3.5 post-radiation, 42 and 48 circRNAs were found to be significantly upregulated and downregulated, respectively, compared to the control (p≤0.05, Fold Change ≥2). Further, the altered expression of 10 circRNAs (chr18: 35610871-35613502+, chr15: 95864225-95894541+, chr3: 96041338-96042928-, chr5: 64096979-64108263+, chr19: 16705875-16710941-, chr5: 134491893-134500149-, chr19: 42562552-42564341+, chr5: 32640331-32664400+, chr3: 72958113-72960367- and chr8: 79343654-79372364-) were verified by RT-PCR. Compared the miRNA-targeted mRNAs with our mRNAs sequencing data, we found 14 upregulated circRNA-targeted mRNAs were also unregulated and 22 downregulated circRNAs-targeted mRNAs were also downregulated. Gene ontology and KEGG pathway analyses indicated the predicted genes were mainly involved in the MAPK signaling pathway. Conclusions: This study reveals that expression of circRNAs was altered in the jejuna of mice post-irradiation and provides a resource for the study of circRNAs in radiation-induced intestinal injury and repair.

Analysis of changes to lncRNAs and their target mRNAs in murine jejunum after radiation treatment

“…Although our laboratory has found that mesenchymal stem cell and 6‐shogaol can improve animal survival and intestinal function following irradiation injury, we still need to understand more about the effects of radiation in order to solve the problem of radiation‐induced intestinal damage from the effect itself. Therefore, our previous work has analysed the regulatory network of circRNAs in radiation‐induced intestinal injury, as well as some important responses of mRNAs . In this article, we focus on the role of lncRNAs.…”

Expression Profiles of CircRNA and mRNA in Lacrimal Glands of AQP5–/– Mice With Primary Dry Eye

“…The BioAnalyzer 2100 system was used for ensuring quality control and library quantification. Then, 10-pM libraries were denatured into single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ , and clustered, and finally, 150 cycles of sequencing were performed on the Illumina HiSeq sequencer according to the manufacturer’s instructions ( Lu et al, 2018 ).…”

“…According to the manufacturer's instructions (Lu et al, 2018), Trizol (Invitrogen, Carlsbad, CA, United States) was used to extract RNA from the lacrimal glands of both AQP5 +/+ and AQP5 −/− mice, and the optical density (OD 260/280 ) value was determined by using a NanoDrop ND-2000 instrument (Thermo Fisher Scientific, Waltham, MA, United States) to measure the RNA concentration. RNA integrity was determined by agarose gel electrophoresis.…”

Analysis of Circular RNA Expression Profile in HEK 293T Cells Exposed to Ionizing Radiation

“…RNA was isolated from the control group and the irradiation group using TRIzol reagent (Invitrogen, Carlsbad, California) according to the manufacturer’s instructions. 9 Each of the RNA sample concentration was measured using a NanoDrop ND-2000 instrument (Thermo, Waltham, Massachusetts). Besides, denaturing agarose gel electrophoresis was used to assess the integrity of RNA.…”

“…Ten pM libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 150 cycles on Illumina HiSeq Sequencer according to the manufacturer’s instructions. 9…”