Duan Liju, Jiang Hongying, Liu Jifeng, Liu Yilin, Ma Tengfei, Xie Yike, Wang Ling, Cheng Juan, Zou Jian, Wu Jiang, Liu Shixi, Gao Mingzhong, Li Weimin, Xie Heping
Wangjiang Hospital, Sichuan University, Chengdu, China.
Department of Rehabilitation Medicine Center, West China Hospital, Sichuan University, Chengdu, China.
Front Genet. 2021 Sep 16;12:698046. doi: 10.3389/fgene.2021.698046. eCollection 2021.
Prior studies have shown that the proliferation of V79 lung fibroblast cells could be inhibited by low background radiation (LBR) in deep underground laboratory (DUGL). In the current study, we revealed further molecular changes by performing whole transcriptome analysis on the expression profiles of long non-coding RNA (lncRNA), messenger RNA (mRNA), circular RNA (circRNA) and microRNA (miRNA) in V79 cells cultured for two days in a DUGL. Whole transcriptome analysis including lncRNA, mRNAs, circ RNA and miRNA was performed in V79 cells cultured for two days in DUGL and above ground laboratory (AGL), respectively. The differentially expressed (DE) lncRNA, mRNA, circRNA, and miRNA in V79 cells were identified by the comparison between DUGL and AGL groups. Quantitative real-time polymerase chain reaction(qRT-PCR)was conducted to verify the selected RNA sequencings. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was analyzed for the DE mRNAs which enabled to predict target genes of lncRNA and host genes of circRNA. With |log(Fold-change)| ≥ 1.0 and < 0.05, a total of 1257 mRNAs (353 mRNAs up-regulated, 904 mRNAs down-regulated), 866 lncRNAs (145 lncRNAs up-regulated, 721 lncRNAs down-regulated), and 474 circRNAs (247 circRNAs up-regulated, 227 circRNAs down-regulated) were significantly altered between the two groups. There was no significant difference in miRNA between the two groups. The altered RNA profiles were mainly discovered in lncRNAs, mRNAs and circRNAs. DE RNAs were involved in many pathways including ECM-RI, PI3K-Akt signaling, RNA transport and the cell cycle under the LBR stress of the deep underground environment. Taken together, these results suggest that the LBR in the DUGL could induce transcriptional repression, thus reducing metabolic process and reprogramming the overall gene expression profile in V79 cells.
先前的研究表明，在深地实验室（DUGL）中，低本底辐射（LBR）可抑制V79肺成纤维细胞的增殖。在本研究中，我们通过对在DUGL中培养两天的V79细胞的长链非编码RNA（lncRNA）、信使RNA（mRNA）、环状RNA（circRNA）和微小RNA（miRNA）的表达谱进行全转录组分析，进一步揭示了分子变化。分别在DUGL和地面实验室（AGL）中对培养两天的V79细胞进行了包括lncRNA、mRNA、circRNA和miRNA的全转录组分析。通过比较DUGL组和AGL组，鉴定出V79细胞中差异表达（DE）的lncRNA、mRNA、circRNA和miRNA。进行定量实时聚合酶链反应（qRT-PCR）以验证所选的RNA测序结果。然后，对DE mRNA进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路分析，从而能够预测lncRNA的靶基因和circRNA的宿主基因。当|log（倍数变化）|≥1.0且P<0.05时，两组之间共有1257个mRNA（353个mRNA上调，904个mRNA下调）、866个lncRNA（145个lncRNA上调，721个lncRNA下调）和474个circRNA（247个circRNA上调，227个circRNA下调）发生了显著变化。两组之间的miRNA没有显著差异。改变的RNA谱主要在lncRNA、mRNA和circRNA中发现。在深地环境的LBR应激下，DE RNA参与了许多途径，包括ECM-RI、PI3K-Akt信号传导、RNA转运和细胞周期。综上所述，这些结果表明DUGL中的LBR可诱导转录抑制，从而减少代谢过程并重新编程V79细胞中的整体基因表达谱。
